WO2024188214A1 - Tricyclic compound, and preparation and application thereof - Google Patents

Abstract

A type of tricyclic compound having the structure shown in formula (I), a preparation method therefor, and a use thereof in modulating CDK2 kinase activity and/or preventing and/or treating CDK2-related diseases.

Description

A tricyclic compound and its preparation and application Technical Field

The present invention relates to the field of medical technology, and in particular to a class of tricyclic compounds used as CDK2 kinase inhibitors, and pharmaceutically acceptable salts thereof, to pharmaceutical compositions comprising such compounds and salts, and to uses thereof, and to applications thereof in regulating CDK2 kinase activity or preventing and/or treating CDK2-related diseases.

Background Art

Cyclin-dependent kinases (CDKs) are a family of serine/threonine kinases, important cellular enzymes that perform essential functions in regulating cell division and proliferation.

Heterodimerization of CDKs and regulatory subunits (Cyclins) allows their full activation and regulation of key cellular processes including cell cycle progression and cell division. Uncontrolled proliferation is a hallmark of cancer cells. Aberrant regulation of CDK activity is associated with aberrant regulation of the cell cycle and is detected in nearly all forms of human cancer.

CDK2 plays a crucial role in the G1/S transition and S phase progression. CDK2 and cyclin E (CCNE) complex phosphorylates retinoblastoma pocket protein family members (p107, p130, pRb), releases E2F transcription factor, and promotes the transition from G1 phase to S phase. This in turn activates CDK2/cyclin A to phosphorylate endogenous substrates, thereby allowing DNA synthesis, replication, and centrosome duplication. It has been reported that the CDK2 pathway affects tumorigenesis mainly through the amplification and/or overexpression of CCNE1 and the mutational inactivation of endogenous inhibitors of CDK2 (such as p27).

CCNE1 copy number gain and overexpression have been demonstrated in ovarian, gastric, endometrial, breast, and other tumors and are associated with poor prognosis in these tumors. CCNE1 amplification and/or overexpression have also been reported to lead to trastuzumab resistance in HER2+ breast cancer and resistance to CDK4/6 inhibitors in estrogen receptor-positive breast cancer. Inhibitors targeting CDK2 have been shown to induce apoptosis and inhibit tumor proliferation.

However, due to the high similarity of the ATP-binding domains of CDKs family kinases, there is currently a lack of selective inhibitors for CDK2. Most small molecules can often inhibit multiple CDK kinase subtypes and even other non-CDK family kinases at the same time. Such off-target effects not only bring about potent cell killing or inhibition, but also bring about significant side effects. Therefore, the development of new selective CDK2 kinase inhibitors has broad application prospects.

Summary of the invention

The object of the present invention is to provide a compound represented by formula (I), a preparation method thereof and use thereof in CDK2 kinase inhibitor drugs.

In the first aspect of the present invention, a compound is provided, wherein the compound is a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof,

in:

Ring A is a saturated or partially saturated 5-6 membered heterocycloalkyl group containing 1 or 2 heteroatoms selected from N, O or S;

Ring B is selected from the group consisting of C _5-6 aryl, 5-6 membered heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S;

Each _Ra , _Rc , and _Rd is independently selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, cyano, =O, _C1-4 alkyl, C1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated _C1-4 alkyl, deuterated _C1-4 alkyl, halogenated _C1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, _C1-4 alkoxy, halogenated _C1-4 alkoxy, and _C3-4 cycloalkyl _;

is a single bond or a double bond;

p, q, t, w are each independently selected from the following group: 0, 1, 2, 3, 4;

s is selected from the following group: 1, 2;

R ₁ is selected from the following group: -CONR ₂ R ₃ , -COOR ₂ , 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is optionally substituted with 0, 1, 2, 3, 4, 5 or 6 R ₄ ;

_X1 is selected from the group consisting of _CH2 , O; or _X1 is absent, such that Four-membered cyclobutyl

X ₂ is selected from the group consisting of O, -CH ₂ O-, NH, -CH ₂ NH-;

R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _{3-12 cycloalkyl} ), -L-(3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, amino, C _1-4 alkyl, C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl, halogenated C _1-4 alkoxy, -S(O) ₂ -C _1-4 alkyl,

Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy;

L is a bond or a C _1-6 alkylene group, wherein the C _1-6 alkylene group is optionally substituted by 0, 1, 2 or 3 substituents selected from the group consisting of deuterium, =O, hydroxyl, halogen, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo-C _1-4 alkyl, halo-C _1-4 alkoxy;

Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy;

R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl;

Each R ₇ is independently selected from the group consisting of hydrogen, deuterium, halogen, C _1-4 alkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl.

In another preferred embodiment, Select from the following group:

Each of _Ra1 , _Rb1 , _Rc1 , and _Rd1 is independently selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, cyano, =O, _C1-4 alkyl, _C1-4 heteroalkyl containing 1, 2 or 3 heteroatoms _selected from N, O or S, halogenated _C1-4 alkyl, deuterated C1-4 alkyl, halogenated _C1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, _C1-4 alkoxy, halogenated _C1-4 alkoxy, and _C3-4 cycloalkyl;

n, m, and r are each independently selected from the following group: 0, 1, 2, and 3.

In another preferred embodiment, the compound has a structure selected from the following group:

Among them, ring A, ring B, R ₁ , _Ra , R _c , R _d , p, q, t are as defined in claim 1.

In another preferred embodiment, the compound has a structure selected from the following group:

Among them, ring A, ring B, _Ra , _Rc , _Rd , p, q, t as defined in claim 1;

R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _{3-12 cycloalkyl} ), -L-(3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, _{cyano, =O, C 1-4 alkyl} , C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl _{, halogenated C 1-4 alkoxy} ,

Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy;

L as defined in claim 1;

Ring C is selected from the following group: 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ;

Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy;

R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl.

In another preferred embodiment, the compound has a structure selected from the following group:

Among them, ring A, ring B, _Ra , _Rc , _Rd , p, q, t as defined in claim 1;

R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _{3-12 cycloalkyl} ), -L-(3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, _{cyano, =O, C 1-4 alkyl} , C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl _{, halogenated C 1-4 alkoxy} ,

Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, the 3-7 membered heterocycloalkyl being optionally substituted by 0, 1, 2, 3 or 4 Substituted by a substituent selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, C _1-4 alkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl, halogenated C _1-4 alkoxy;

L as defined in claim 1;

Ring C is selected from the following group: 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ;

Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy;

R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl.

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, Preferably

In another preferred embodiment, the compound has a structure selected from the following group:

in,

R ₁ is selected from the following group: -CONR ₂ R ₃ , 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ;

R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _{3-12 cycloalkyl} ), -L-(3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, _{cyano, =O, C 1-4 alkyl} , C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl _{, halogenated C 1-4 alkoxy} ,

Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy;

L as defined in claim 1;

Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl C _1-6 alkyl, C _1-6 alkoxy, C 1-6 alkoxy halogenated C 1-6 alkyl, C _1-6 alkoxy halogenated C 1-6 alkoxy alkyl, C 1-6 alkoxy halogenated C 1-6 alkoxy alkyl, C 1-6 alkoxy halogenated C _1-4 alkyl, C _1-4 alkoxy halogenated C _1-4 alkoxy alkyl, C _1-6 ...

R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl;

R _a1 is selected from the group consisting of hydrogen, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, deuterated C 1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, ₂ or 3 heteroatoms selected from N, O or S, and C _3-4 cycloalkyl;

Each R _c1 is independently selected from the following group: hydrogen, deuterium, halogen, hydroxyl, cyano, =O, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-4 alkoxy, halogenated C _1-4 alkoxy;

Each R _d1 is independently selected from the group consisting of hydrogen, deuterium, halogen, cyano, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-4 alkoxy, halogenated C _1-4 alkoxy;

m and r are each independently selected from the following group: 0, 1, 2, 3.

In another preferred embodiment, R _a1 is selected from the following group: C _1-4 alkyl, deuterated C _1-4 alkyl.

In another preferred embodiment, each R _c1 is independently selected from the following group: hydrogen, halogen, hydroxyl, =0.

In another preferred embodiment, each R _d1 is independently selected from the group consisting of hydrogen, C _1-4 alkyl, and halogenated C _1-4 alkyl.

In another preferred embodiment, the compound has a structure selected from the following group:

in,

R ₁ is selected from the following group: -CONR ₂ R ₃ , 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ;

R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _{3-12 cycloalkyl} ), -L-(3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, _{cyano, =O, C 1-4 alkyl} , C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl _{, halogenated C 1-4 alkoxy} ,

Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy;

L as defined in claim 1;

Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy;

R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl;

R _a1 is selected from the group consisting of hydrogen, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, deuterated C 1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, ₂ or 3 heteroatoms selected from N, O or S;

Each R _c1 is independently selected from the following group: hydrogen, deuterium, halogen, hydroxyl, cyano, =O, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-4 alkoxy, halogenated C _1-4 alkoxy;

Each _Rd1 is independently selected from the group consisting of hydrogen, deuterium, halogen, cyano, _C1-4 alkyl, _C1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated _C1-4 alkyl, A halogenated C _1-4 heteroalkyl group, a C _1-4 alkoxy group, or a halogenated C _1-4 alkoxy group selected from a heteroatom of N, O or S;

m and r are each independently selected from the following group: 0, 1, 2, 3.

In another preferred embodiment, R ₁ is -CONR ₂ R ₃ .

In another preferred embodiment, R ₁ is a 5-6 membered monocyclic heteroaryl group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S.

In another preferred embodiment, R ₁ is a 3-6 membered heterocycloalkyl group containing 1, 2, 3 or 4 heteroatoms selected from N, O or S.

In another preferred embodiment, R _a1 is selected from the following group: C _1-4 alkyl, deuterated C _1-4 alkyl.

In another preferred embodiment, _Ra1 is -CH ₃ .

In another preferred embodiment, _Ra1 is -CD ₃ .

In another preferred embodiment, R _c1 is selected from the following group: hydrogen, halogen, hydroxyl, =O.

In another preferred embodiment, R _c1 is hydrogen.

In another preferred embodiment, R _c1 is halogen.

In another preferred embodiment, R _c1 is F.

In another preferred embodiment, R _c1 is ＝O.

In another preferred embodiment, R _d1 is selected from the group consisting of hydrogen, C _1-4 alkyl, and halogenated C _1-4 alkyl.

In another preferred embodiment, R _d1 is hydrogen.

In another preferred embodiment, R _d1 is -CH ₃ .

In another preferred embodiment, R _d1 is -CF ₃ .

In another preferred embodiment, R _d1 is -CHF ₂ .

In another preferred embodiment, m is 0.

In another preferred embodiment, m is 1.

In another preferred embodiment, m is 2.

In another preferred embodiment, r is 0.

In another preferred embodiment, r is 1.

In another preferred embodiment, the compound has a structure selected from the following group:

in,

R ₁ is selected from the following group: -CONR ₂ R ₃ , 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ;

R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _{3-12 cycloalkyl} ), -L-(3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, _{cyano, =O, C 1-4 alkyl} , C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl _{, halogenated C 1-4 alkoxy} ,

Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy;

L as defined in claim 1;

Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy;

R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl;

Each R _d1 is independently selected from the group consisting of hydrogen, deuterium, halogen, cyano, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-4 alkoxy, halogenated C _1-4 alkoxy;

r is selected from the following group: 0, 1.

In another preferred embodiment, the structure Select from the following group:

In another preferred embodiment, each R _d1 is independently selected from the group consisting of hydrogen, C _1-4 alkyl, and halogenated C _1-4 alkyl.

In another preferred embodiment, the compound is selected from the following group:

The second aspect of the present invention provides a pharmaceutical composition comprising a preventive and/or therapeutically effective amount of one or more compounds described in the first aspect of the present invention and a pharmaceutically acceptable carrier.

The third aspect of the present invention provides a use of the compound described in the first aspect of the present invention for preparing a drug for regulating CDK activity or for preventing and/or treating CDK-related diseases.

In another preferred embodiment, the CDK is CDK2.

In another preferred embodiment, the CDK-related disease is a disease with high expression of CDK2.

In another preferred embodiment, the CDK-related disease is a disease caused by abnormal expression or amplification of Cyclin E.

In another preferred embodiment, the CDK-related disease is cancer.

In another preferred embodiment, the cancer is a cancer caused by abnormal expression or amplification of CyclinE.

In another preferred embodiment, the cancer is selected from the following group: breast cancer, ovarian cancer, bladder cancer, prostate cancer, melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer (including HCC), pancreatic cancer, colorectal cancer, lung cancer (including NSCLC, SCLC, squamous cell carcinoma or adenocarcinoma), renal cancer (including RCC), skin cancer, glioblastoma, glioblastoma, mantle cell lymphoma (MCL), chronic myeloid leukemia (CML), acute myeloid leukemia (AML), neuroblastoma, sarcoma, liposarcoma, chondroma, osteoma, osteosarcoma, seminoma, testicular tumor, uterine cancer, head and neck cancer, multiple myeloma, malignant lymphoma, polycythemia vera, leukemia, thyroid cancer, ureteral tumor, bladder tumor, gallbladder cancer, bile duct cancer, choriocarcinoma, pediatric tumor.

The fourth aspect of the present invention provides a use of the compound described in the first aspect of the present invention for preparing a medicament for preventing and/or treating a cell cycle protein-related disease.

In another preferred embodiment, the cell cycle protein is cyclin E.

In another preferred embodiment, the cyclin E is selected from the following group: cyclin E1 and cyclin E2.

In another preferred embodiment, the cell cycle protein-related disease is cancer.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

DETAILED DESCRIPTION

After long-term and in-depth research, the inventor unexpectedly prepared a class of tricyclic compounds with excellent CDK kinase inhibitory activity. Specifically, through structural optimization, the inventor obtained a class of compounds with excellent CDK2 selective inhibitory activity. On this basis, the inventor completed the present invention.

the term

In the present invention, unless otherwise specified, the terms used have the general meanings well known to those skilled in the art.

In the present invention, the term "halogen" refers to F, Cl, Br or I.

In the present invention, "C _1-6 alkyl" refers to a straight or branched alkyl group including 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, or the like. The term "C _1-4 alkyl" has a similar meaning.

In the present invention, the term "C _3-12 cycloalkyl" refers to a cyclic alkyl group having 3-12 carbon atoms in the ring, wherein the ring includes C _3-6 monocycloalkyl, C _5-12 bicycloalkyl, C _5-10 bridged ring group, C _6-10 spirocyclic group, including but _not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,

In the present invention, the term "C _1-4 alkoxy" refers to a straight or branched alkoxy group having 1 to 4 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy and butoxy.

In the present invention, the term "C _1-6 alkylene" refers to a _straight or branched _alkylene structure including 1 to _{6 carbon} atoms, for _{example , -CH2- , -CH2CH2- , -CH2CH2CH2- , -CH2CH2CH2CH2- , -CH2CH2CH2CH2CH2- , -CH2CH2CH2CH2CH2- , -CH2CH2CH2CH2CH2CH2- , -CH2CH2CH2CH2CH2CH2- .}

In the present invention, the term "C _1-6 heteroalkyl" refers to a straight or branched alkyl group having 1 to 6 carbon atoms containing 1 to 4 heteroatoms selected from N, O or S, including but not limited to methoxymethyl, ethoxymethyl, propoxymethyl, isopropoxymethyl, butoxymethyl, -CH ₂ OH, -CH ₂ NH ₂ , -CH ₂ NHCH ₃ , -CH ₂ N(CH ₃ ) ₂ , -CH ₂ CH ₂ OCH ₃ , -CH ₂ CH ₂ NHCH ₃ , -CH ₂ CH ₂ N(CH ₃ ) ₂ , etc. The similar term "C _1-4 heteroalkyl" has a similar meaning.

In the present invention, the term "deuterated C _1-4 alkyl" refers to an alkyl group having 1 to 4 carbon atoms in which one, more or all hydrogen atoms on the carbon atoms of a linear or branched chain are optionally substituted with deuterium, for example, -CD ₃ , -CH ₂ CD ₃ , -CH(CH ₃ )CD ₃ and the like.

In the present invention, the term "heterocycloalkyl" refers to a saturated or partially saturated 3-12-membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O, and S, wherein the heterocycle comprises a 3-6-membered monocyclic heteroalkyl, a 5-12-membered bicyclic heteroalkyl, a 5-10-membered heterobridged ring group, and a 6-10-membered heterospirocyclic group, including (but not limited to) the following groups:

In the present invention, the term "aromatic ring" or "aryl group" has the same meaning, preferably "C _5-6 aryl group". The term "C _5-6 aryl group" refers to an aromatic ring group having 5 to 6 carbon atoms without heteroatoms in the ring, such as phenyl and the like.

In the present invention, the term "aromatic heterocycle" or "heteroaryl" has the same meaning and refers to a heteroaromatic group containing one to multiple heteroatoms. For example, "5-6 membered heteroaryl" refers to an aromatic heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen and 1 to 5 carbon atoms. Non-limiting examples include: furanyl, thienyl, pyridyl, pyrazolyl, thiazolyl, isothiazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. The heteroaryl group can be optionally substituted or unsubstituted.

In the present invention, the term "monocyclic heteroaryl" refers to a heteroaromatic group having only one ring and containing one or more heteroatoms. For example, "5-6 membered monocyclic heteroaryl" refers to a monocyclic aromatic heterocycle containing 1 to 4 heteroatoms selected from O, S and N and 1 to 5 carbon atoms. Non-limiting examples include: furanyl, thienyl, pyridyl, pyrazolyl, thiazolyl, isothiazolyl, pyrrolyl, N-alkylpyrrolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl group may be optionally substituted or unsubstituted.

In the present invention, the term "bicyclic heteroaryl" refers to a heteroaromatic group having two rings containing one or more heteroatoms. For example, "7-10 membered bicyclic heteroaryl" refers to a bicyclic aromatic heterocycle containing 1 to 4 heteroatoms selected from O, S and N and 3 to 9 carbon atoms. Non-limiting examples include:
etc. The heteroaryl group may be optionally substituted or unsubstituted.

In the present invention, the term "halo" means substituted with halogen.

In the present invention, the term "substituted" refers to one or more hydrogen atoms on a specific group being replaced by a specific substituent. The specific substituent is a substituent described above or a substituent appearing in each embodiment. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable position of the group, and the substituent may be the same or different at each position. It should be understood by those skilled in the art that the combination of substituents contemplated by the present invention is a combination that is stable or chemically feasible. The substituents include, for example (but not limited to): deuterium, halogen, hydroxyl, cyano, amino, =O, C _1-6 alkyl, C _1-6 alkoxy, C _3-10 cycloalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _3-10 halocycloalkyl, _3-10 heterocyclic group, aryl, heteroaryl, C2-C10 acyl, C2-C10 ester, etc.

In the present invention, the term 1-4 means 1, 2, 3 or 4. Other similar terms have similar meanings.

In the present invention, "-CO-" means -(C=O)-.

In the present invention, "-SO ₂ -" means

It should be understood that when a group is present in multiple different positions of a compound at the same time, its definition at each position is independent of each other and may be the same or different. That is, the term "selected from the following group:" has the same meaning as the term "independently selected from the following group:".

Compound

The present invention provides a compound, which is a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof,

Among them, ring A, ring B, R ₁ , R ₇ , _Ra , R _c , R _d , p, q, t, _Xi , X ₂ , s and w are as defined above.

In another preferred embodiment, in the compound, ring A, ring B, Any one of R ₁ , R ₇ , _Ra , R _c , R _d , p, q, t, X ₁ , X ₂ , s, and w is independently a corresponding specific group in the specific compound.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Acids suitable for forming salts include but are not limited to Limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, glutamic acid.

Another preferred salt is a salt of the compound of the present invention and a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

The term "solvate" refers to a complex in which the compound of the present invention is coordinated with solvent molecules to form a specific ratio.

In addition, the compounds of the present invention also include prodrugs of the compounds shown in formula (I). The term "prodrug" includes a compound that may be biologically active or inactive, and when taken by an appropriate method, it undergoes metabolism or chemical reactions in the human body and is converted into a class of compounds of formula (I), or a salt or solution composed of a compound of formula (I). The prodrug includes (but is not limited to) carboxylate, carbonate, phosphate, nitrate, sulfate, sulfone, sulfoxide, amino compound, carbamate, azo compound, phosphoramide, glucoside, ether, acetal and the like forms of the compound.

It should be understood that the specific preparation method of the compound of formula (I) of the present invention does not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combination can be easily performed by a person skilled in the art to which the present invention belongs.

Typically, the compounds of the present invention can be prepared by the process described in the examples, wherein the raw materials and reagents used can be purchased from commercial sources unless otherwise specified.

Pharmaceutical compositions and methods of administration

The present invention also provides a pharmaceutical composition comprising a preventive and/or therapeutically effective amount of one or more compounds of the present invention and a pharmaceutically acceptable carrier.

Since the compounds of the present invention have excellent anti-tumor activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate tumor-related diseases.

The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-1000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The pharmaceutical composition is in the form of injection, capsule, tablet, pill, powder or granule.

There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may include physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and solutions for reconstitution into sterile injectable solutions or Sterile powders for dispersion. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as anti-tumor drugs).

The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

Compared with the prior art, the present invention has the following main advantages:

1) The compound has excellent CDK2 inhibitory activity;

2) The compound has excellent CDK2 selective inhibitory activity;

3) The compound has a selective inhibitory effect on cancer caused by abnormal expression or amplification of CyclinE;

4) The compound has excellent pharmacokinetic properties.

The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are generally carried out according to conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

Example Synthesis Route

Unless otherwise stated, the absolute stereochemistry of all chiral atoms is defined as follows. The "&" symbol in the structural formula indicates that the compound is a cis racemate, the "or" symbol indicates that the compound is a single-configuration compound, but the absolute configuration is unknown, the "abs" symbol indicates that the absolute configuration of the compound is the structure shown, and the "rac" symbol indicates that the compound is a racemate.

Synthesis of intermediates

Synthesis of intermediate int1:

1. Synthesis of int1-2

Dissolve int1-1 (10 g, 64.0 mmol) in 150 mL of methanol, add NaBH ₄ (2.44 g, 64.0 mmol) in batches under nitrogen protection and ice-water bath cooling, then heat the reaction solution to room temperature and stir for 2 hours. After the reaction is completed, slowly add 1 M dilute hydrochloric acid solution under ice-water bath cooling to quench the reaction, concentrate under reduced pressure to evaporate the solvent methanol, then dilute with water, extract with dichloromethane, separate the organic phase, dry with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 8.27 g of crude product. Ms[M+Na] ⁺ 181.1

2. Synthesis of int1-3

Dissolve int1-2 (8.27 g, 52.33 mmol) in 100 mL of ethanol, then add imidazole (10.67 g, 157.0 mmol), slowly dropwise add TBDPSCl (15.83 g, 57.58 mmol) under nitrogen protection in an ice-water bath, and after the addition is complete, warm the reaction solution to room temperature and stir for 16 h. After the reaction is completed, evaporate the solvent ethanol under reduced pressure, then dilute the residue with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and column chromatograph to obtain 19.86 g of the product. Ms[M+H] ⁺ : 397.2. ¹ HNMR (400MHz, CDCl ₃ ) δ7.73-7.58(m,4H),7.43-7.33(m,6H),4.31-4.20(m,1H),4.18-4.03(m,2H),2.71-2.55(m,1H),2.19-2.08(m,1H),2.01(d,J ＝5.6Hz,2H),1.86-1.70(m,2H),1.64-1.53(m,1H),1.25(t,J＝7.1Hz,3H),1.05(s,9H).

3. Synthesis of int1-4

Under nitrogen protection, n-butyl lithium (88 mL, 2.5 M in hexane) was added to 500 mL of anhydrous tetrahydrofuran, the system was cooled to -65 ° C, and then acetonitrile (9.2 g, 224.8 mmol) was slowly added dropwise. After the addition was complete, the temperature was maintained and stirred for 1 hour. Then, an anhydrous tetrahydrofuran solution (100 mL) of int1-3 (44.7 g, 112.4 mmol) was slowly added dropwise, and the temperature was maintained and stirred for 2 hours. After the reaction was completed, water was added dropwise at -65 ° C to quench the reaction, and then stirred and heated to room temperature. The reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 49.0 g of crude product. Ms [M + H] ⁺ : 392.2

4. Synthesis of int1-5

Dissolve tert-butylhydrazine hydrochloride (9.35 g, 75.0 mmol) and triethylamine (10.43 mL, 75.0 mmol) in 150 mL of ethanol, stir for 0.5 hours, then add ethanol solution (50 mL) of int1-4 (19.6 g, 50.0 mmol), and then heat the reaction system to 75 ° C and stir for 16 hours. After the reaction is completed, The reaction solution was cooled to room temperature, concentrated, diluted with water, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified to obtain 19.1 g. Ms[M+H] ⁺ : 462.3 ¹ H NMR (400MHz, CDCl ₃ )δ7.70-7.66(m,4H),7.40-7.31(m,6H),5.46(s,1H),5.19(s,1H),4.46-4.27(m,1H),3.55-3.27(m,2H),2.90-2.81(m,1H),2.29-2.04(m,1H),1.93-1.64(m,4H),1.57(s,9H),1.06(s,9H).

5. Synthesis of int1-6

Dissolve int1-5 (19.12 g, 41.0 mmol) in 160 mL of acetonitrile, add benzyl chloroformate (11.7 mL, 82.0 mmol) dropwise under ice-water cooling, warm to room temperature and stir for 2 h, then add NaHCO ₃ (11.02 g, 130.0 mmol) and stir at room temperature for 16 h. After the reaction is completed, filter the reaction solution, wash the filter cake with ethyl acetate several times, concentrate the filtrate under reduced pressure, dilute with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate under reduced pressure and purify to obtain 21.2 g. Ms[M+H] ⁺ : 596.4

6. Synthesis of int1-7

Int1-6 (21.0 g, 35.0 mmol) was dissolved in 150 mL of anhydrous tetrahydrofuran under stirring at room temperature, and glacial acetic acid (4.2 mL, 70.0 mmol) and tetrabutylammonium fluoride (35.0 mL, 1 M in THF) were added in sequence. The mixture was heated to 40°C and stirred for 23 hours under a nitrogen atmosphere. After the reaction, the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the residue was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified to obtain 4.41 g. Ms[M+H] ⁺ : 358.2. ¹ H NMR (400MHz, DMSO-d6) δ9.06 (s, 1H), 7.59-7.22 (m, 5H), 5.92 (s, 1H), 5.12 (s, 2H), 4.57 (d, J = 4.4Hz, 1H), 4.22-4.09 (m, 1H), 2.99-2.78 (m, 1H), 2.23 -2.14(m,1H),1.90-1.79(m,1H),1.77-1.64(m,3H),1.47(s,9H).

7. Synthesis of int1-8

Dissolve int1-7 (4.41 g, 12.35 mmol) in 80 mL of dichloromethane, add p-nitrophenyl chloroformate (3.73 g, 18.53 mmol), pyridine (2.93 g, 37.05 mmol), DMAP (149 mg, 1.24 mmol) in sequence, and stir at room temperature for 16 h. After the reaction, the reaction solution was washed with saturated brine for several times, dried over anhydrous sodium sulfate, concentrated, and purified to obtain 4.34 g of the product. Ms[M+H] ⁺ : 523.2

8. Synthesis of int1-9

Dissolve int1-8 (4.34 g, 8.31 mmol) in 80 mL of tetrahydrofuran, add isopropylamine (1.07 mL, 12.47 mmol) and diisopropylethylamine (4.33 mL, 24.93 mmol) in turn, and stir at room temperature overnight. After the reaction is completed, dilute the reaction system with water, extract with ethyl acetate, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify to obtain 1.17 g of the product. Ms[M+H] ⁺ : 443.3

9. Synthesis of int1, int1A, int1B

Int1-9 (1.07 g, 2.42 mmol) was dissolved in a mixed solvent of 10 mL methanol and 10 mL ethyl acetate, and then Pd/C (214 mg, 20 mol%) was added and stirred at room temperature for 5 h under a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and purified to obtain 640 mg of int1. The peak time of int1A (300 mg) was 28.878 min using chiral preparative chromatography, and the peak time of int1B (291 mg) was 33.753 min. The chromatographic column used was Phenomenex Cellulose-4, mobile phase is n-hexane:isopropanol=90:10, column temperature is 30°C.

Synthesis of intermediates int1C and int1D:

1. Synthesis of int1C-1

The following method was used for chiral separation of int1-7: Agilent HPLC separation, the preparation and separation conditions were as follows: Chromatographic column: Phenomenex 5μm Cellulose-4; mobile phase: n-hexane:isopropanol = 90:10; flow rate: 1mL/min, detection wavelength: 220nm; column temperature: 30℃, solvent: isopropanol, collect the sample with peak time at 24.6min, concentrate and freeze-dry to obtain the product int1C-1, the peak time of another configuration is 21.8min.

2. Synthesis of int1C

Dissolve intC1-1 (2.2 g, 6.2 mmol) in 40 mL of dichloromethane, add p-nitrophenyl chloroformate (1.86 g, 9.26 mmol), pyridine (1.47 g, 18.5 mmol), DMAP (74.5 mg) in sequence, and stir at room temperature for 16 h. After the reaction is completed, the reaction solution is washed with saturated brine for several times, dried over anhydrous sodium sulfate, concentrated, and purified to obtain 2.21 g of the product.

3. Synthesis of int1D-1

In a 50mL single-mouth bottle, int1C (2.0g, 1.0eq) was added, and then formic acid (40mL) was added at room temperature, and the reaction was carried out at 80°C for 2h. After the reaction was completed by TLC monitoring, the pH was adjusted to near neutral with saturated sodium bicarbonate, and then extracted with ethyl acetate (100ml x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1.2g of the product.

4. Synthesis of int1D

In a 250mL single-necked bottle, int1D-1 (4.66g, 1.0eq), dichloromethane (100mL), and DHP (1.68g) were added. 344mg of p-toluenesulfonic acid was added dropwise at room temperature. The mixture was reacted at 40°C overnight, diluted with water, extracted with dichloromethane (100ml x 3), and the organic phases were combined. The organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography was used to obtain 5.23g of the product.

Synthesis of intermediate int2:

1. Synthesis of int2-2

Add tetrahydrofuran (240 mL) to a 1000 mL three-necked flask, add NaH (60%) (4.3 g 2 eq) under nitrogen protection, stir for 5 min, then slowly add diethyl malonate (21.27 g 2.5 eq) dropwise After the addition, stir at room temperature for 30 minutes, then dropwise add int2-1 (10 g, 1 eq)/tetrahydrofuran (120 ml), and keep at 80°C for 16 hours. After the reaction is completed, add 400 ml of saturated ammonium chloride to the reaction solution, extract with ethyl acetate (100 ml x 3), combine the organic phases, dry the organic phases over anhydrous sodium sulfate, filter, concentrate, and perform column chromatography to obtain 8.3 g of int2-2.

¹ H NMR (400MHz, CDCl ₃ ) δ8.58 (d, J = 5.8Hz, 1H), 7.00 (d, J = 5.8Hz, 1H), 4.95 (s, 1H), 4.28 (qd, J = 7.1, 1.1Hz, 4H), 3.99 (s, 3H), 1.28 (t, J = 7.2Hz, 6H).

2. Synthesis of int2-3

In a 250 mL single-mouth bottle, int2-2 (8.3 g, 1.0 eq), methanol (83 mL), Pd/C (0.83 g) were added and reacted at 30 °C for 3 h. After the reaction was completed, the mixture was filtered and dried to obtain 7.2 g of int2-3.

3. Synthesis of int2-4

Int2-3 (7.2 g, 1.0 eq), concentrated hydrochloric acid (50 mL), and water (50 mL) were added to a 250 mL three-necked flask, and the mixture was kept at 100 °C for 16 h under nitrogen protection. After the reaction was completed, the mixture was cooled to room temperature, adjusted to pH = 8 with saturated sodium bicarbonate solution, extracted with ethyl acetate (100 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 2.8 g of int2-4.

¹ H NMR (400MHz, DMSO-d6) δ10.60 (s, 1H), 8.01 (d, J = 5.8Hz, 1H), 6.96 (d, J = 5.9Hz, 1H), 3.87 (s, 3H), 3.56(s,2H).

4. Synthesis of int2-5

In a 100 mL single-mouth bottle, int2-4 (1.2 g, 1.0 eq), DMSO (20 ml), DBU (2 g 2 eq), and Zn(OTf) ₂ (0.5 g 0.2 eq) were added, and diphenyl(vinyl)sulfonium trifluoromethanesulfonate (2.36 g 1 eq) was added after stirring at 25°C for 5 min. The mixture was reacted at 25°C for 1.5 h. After the reaction was completed, a saturated sodium chloride solution (60 ml) was added, and the mixture was extracted with ethyl acetate (100 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 600 mg of int2-5.

¹ H NMR (400MHz, CDCl ₃ ) δ8.08 (d, J＝5.9Hz, 1H), 7.82 (q, J＝5.2, 4.1Hz, 1H), 6.72 (d, J＝5.9Hz, 1H), 3.94 (s,3H),1.89–1.83(m,2H),1.80–1.74(m,2H).

5. Synthesis of int2-6

In a 100mL three-necked flask, add DMF (10mL) and int2-5 (600mg 1eq), cool to 0℃ under nitrogen protection, add NaH (60%) (188mg 1.5eq), stir for 5min, then slowly add iodomethane (892mg 2eq) dropwise, stir at room temperature for 2h. After the reaction is completed, add saturated ammonium chloride 40ml to the reaction solution, and then extract with ethyl acetate (30ml x3). After combining the organic phases, dry the organic phases over anhydrous sodium sulfate, filter, and concentrate to obtain 660mg of int2-6.

¹ H NMR (400MHz, CDCl ₃ ) δ8.03 (d, J = 5.8 Hz, 1H), 6.72 (d, J = 5.9 Hz, 1H), 3.93 (s, 3H), 3.55 (s, 3H), 1.86 –1.78(m,2H),1.78–1.62(m,2H).

6. Synthesis of int2-7

In a 100mL three-necked flask, int2-6 (150mg 1eq), dichloromethane (10ml) and boron tribromide (5ml) were added. The temperature was lowered to 0°C under nitrogen protection, and boron tribromide (5ml) was added. The mixture was stirred at 40°C for 8h. After concentration under reduced pressure, the mixture was dissolved in dichloromethane, quenched with 1ml of water, and dried to obtain 300mg of crude int2-7, which was directly used for the next step.

7. Synthesis of int2

Add int2-7 and dichloromethane (10 ml) into a 100 mL three-necked flask and cool down under nitrogen protection. To 0°C, add trifluoromethanesulfonic anhydride (354mg 1.5eq)/dichloromethane (2ml), stir for 5min, and after the reaction is complete, add saturated ammonium chloride 40ml to the reaction solution, followed by extraction with dichloromethane (30ml x3). After combining the organic phases, dry the organic phases over anhydrous sodium sulfate, filter, concentrate, and separate and purify by preparative chromatography to obtain int2 (165mg).

Synthesis of intermediate int2A:

1. Synthesis of int2A-2

Dissolve dimethyl malonate (3.31g 1.2eq) in N,N-dimethylformamide (25ml), heat to 40℃, add NaH (0.82g 1.2eq), stir and react for 0.5h, then add 4,6-dichloro-5-aminopyrimidine (2.25g1eq), slowly heat to 70℃, keep warm and react for 3h, cool to room temperature after the reaction, remove N,N-dimethylformamide by vacuum distillation, add water (20mL), extract with ethyl acetate (100ml x3), combine organic phases, wash with water (25ml x2), combine aqueous phases and adjust pH to 5-6 with concentrated hydrochloric acid, yellow solid precipitates, filter and dry to obtain 1.9g of product. ¹ H NMR (400MHz, DMSO) δ＝10.98(s,1H),8.03(s,1H),4.15(d,J＝7.1,2H),1.18(s,3H).

2. Synthesis of int2A-3

Add int2A-2 (1.6 g) and acetic acid (80 mL) into a 250 mL reactor, add concentrated hydrochloric acid (80 mL) with stirring, then slowly raise the temperature to 100 °C, keep stirring and react for 15 h, then cool the reaction solution to room temperature, spin dry to obtain 1.3 g of a crude product, which is directly used in the next step.

3. Synthesis of int2A-4

In a 100 mL single-mouth bottle, int2A-3 (1.1 g, 1.0 eq), DMSO (20 ml), DBU (3.11 g 4 eq), and Zn(OTf) ₂ (0.786 g 0.4 eq) were added, stirred at 25°C for 5 min, and then diphenyl(vinyl)sulfonium trifluoromethanesulfonate (1.86 g 1 eq) was added, and the mixture was reacted at 25°C for 10 min. After the reaction was completed, saturated sodium chloride solution (60 ml) was added, and then the organic phases were extracted with ethyl acetate (100 ml x 3), and then concentrated under reduced pressure. The organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 400 mg of the product.

3. Synthesis of int2A

Add N,N-dimethylformamide (10 mL), int2A-4 (400 mg 1 eq) to a 100 mL three-necked flask, cool to 0 °C under N ₂ protection, add cesium carbonate (1.37 g 2 eq), stir for 5 min, slowly add iodomethane (405 mg 1.3 eq), stir at room temperature for 0.5 h, and after the reaction is complete, add saturated ammonium chloride 40 ml to the reaction solution, extract with ethyl acetate (30 ml x 3), combine the organic phases, dry the organic phases with anhydrous sodium sulfate, filter, concentrate, and purify by column chromatography to obtain 240 mg of the product. ¹ H NMR (400 MHz, CDCl ₃ ) δ＝8.54 (s, 1H), 3.64 (s, 3H), 1.98 (dd, J＝8.7, 2.0, 4H).

Synthesis of intermediate int2B:

1. Synthesis of int2B-2

Add int2B-1 (10 g, 1.00 eq) and NN-dimethylformamide (100 mL) to a 250 mL flask, stir to dissolve, then add cesium carbonate (19.3 g, 0.9 eq) and iodomethane (11.2 g, 1.2 eq). React at room temperature for 3 to 5 minutes. After 1 h, the reaction was complete, water was added to quench, and the mixture was extracted with ethyl acetate and separated. The organic phase was washed with saturated sodium chloride solution and separated. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain 5 g of the product. Ms[M+H] ⁺ : 167.0

2. Synthesis of int2B-3

In a 100mL flask, int2B-2 (3.25g, 1.00eq) and tert-butyl alcohol (55mL) were added and stirred to dissolve. Then sodium bicarbonate (4.94g, 3.0eq) was added and stirred in an ice-water bath for 10 minutes. Bromine (5.76g, 6.0eq) was slowly added dropwise. The reaction was allowed to react at room temperature for 1 hour until the reaction was complete. Water was added to quench the mixture, and the mixture was extracted with ethyl acetate and separated. The organic phase was washed with saturated sodium chloride solution and separated. The organic phase was dried over anhydrous sodium sulfate, concentrated, and purified on a silica gel column to obtain 1.78g of the product. Ms[M+H] ⁺ : 338.8.

3. Synthesis of int2B-4

In a 100 mL flask, int2B-3 (1.78 g, 1.00 eq) and acetic acid (36 mL) were added and stirred for 10 minutes in an ice-water bath. Zinc powder (1.73 g, 5.0 eq) was slowly added. The reaction was allowed to proceed at room temperature for 15 minutes. After the reaction was complete, the mixture was filtered, the filter cake was rinsed with methanol, the filtrate was concentrated, and the product (0.75 g) was purified by silica gel column. Ms[M+H] ⁺ : 183.0.

4. Synthesis of int2B

In a 50mL flask, int2B-4 (0.75g, 1.00eq), diphenyl (vinyl) sulfonium trifluoromethanesulfonate (1.49g, 1.00eq), and dichloromethane (7.5mL) were added and stirred to dissolve. Then zinc trifluoromethanesulfonate (0.3g, 0.2eq) was added and stirred at room temperature for 3 minutes. A mixture of DBU (1.25g, 2.0eq) in dichloromethane (7.5mL) was added. The reaction was allowed to react at room temperature for 10 minutes until the reaction was complete. Saturated ammonium chloride solution was added to the system, stirred, and separated. Saturated sodium chloride solution was added to the organic phase, washed, separated, and the organic phase was concentrated and purified on a silica gel column to obtain 0.49g of the product. Ms[M+H] ⁺ : 209.0.

Synthesis of intermediate int2C:

1. Synthesis of int2C-1

In a 100mL single-mouth bottle, int2 (760mg, 1.00eq), int1-5 (1.09g, 1.00eq), cesium carbonate (2.31g, 3.00eq), Xantphos (273mg, 0.20eq), palladium acetate (53mg, 0.10eq) and 1,4-dioxane (15mL) were added. The temperature was raised to 100°C in a nitrogen atmosphere for 2h, and the reaction was complete. The temperature was lowered, filtered, and the solid was washed with ethyl acetate. The organic phase filtrate was collected, and the organic phase was washed three times with saturated brine, and then the organic phase was dried over anhydrous sodium sulfate and concentrated. Purification by silica gel column gave 200mg of the product. Ms[M+H] ⁺ : 634.3.

2. Synthesis of int2C-2

In a 50 mL single-mouth bottle, int2C-1 (200 mg, 1 eq) and methanol (5 mL) were added, and concentrated hydrochloric acid (0.25 mL) was added to the solution. The reaction solution was stirred at 40°C for 2 h until the reaction was complete. The reaction solution was concentrated, and the pH was adjusted to neutral with saturated sodium bicarbonate aqueous solution. The solution was extracted with ethyl acetate three times, and the organic phase was dried over anhydrous sodium sulfate. After concentration, 200 mg of the product was obtained. Ms[M+H] ⁺ : 396.2.

3. Synthesis of int2C-3

In a 50 mL single-mouth bottle, int2C-2 (200 mg), N-methylmorpholine (511 mg, 10 eq) and dichloromethane (20 mL) were added. A solution of p-nitrochloroformic acid phenyl ester (204 mg, 2 eq) in dichloromethane (10 mL) was slowly added dropwise to the reaction solution at 0°C. The reaction solution was stirred at 0°C for 2 min until the reaction was complete. Water was added to the reaction solution at 0°C to quench the reaction, separate the layers, extract the aqueous phase with ethyl acetate three times, combine the organic phases, dry with anhydrous sodium sulfate and concentrate. After purification on a thin layer chromatography preparative plate, 120 mg of the product was obtained. Ms[M+H] ⁺ : 561.3.

4. Synthesis of int2C

In a 25 mL single-necked bottle, int2C-3 (110 mg, 1 eq) and formic acid (5 mL) were added. The temperature was raised to 80°C for reaction for 1.5 h, then the temperature was lowered, and the mixture was concentrated and used directly in the next step. Ms [M+H] ⁺ : 505.2.

Synthesis of intermediate int2D:

1. Synthesis of int2D-2

In a 100mL flask, int2B (4g, 1.00eq), int2D-1 (1.8g, 1.00eq), cesium carbonate (8.5g, 3.00eq), Xantphos (1g, 0.20eq), palladium acetate (0.2g, 0.10eq) and 1,4-dioxane (40mL) were added. The mixture was heated to 100°C in a nitrogen atmosphere for 2h and the reaction was complete. The mixture was cooled, filtered, and the solid was washed with ethyl acetate. The organic phase was diluted with a large amount of ethyl acetate, and the organic phase was washed with saturated brine three times. The organic phase was dried over anhydrous sodium sulfate and concentrated. Purification by silica gel column gave 5.17g of the product. Ms[M+H] ⁺ : 634.3.

2. Synthesis of int2D-3

In a 100mL flask, int2D-2 (5g, 1eq) and methanol (50mL) were added. Concentrated hydrochloric acid (1mL) was added to the solution. The reaction solution was stirred at 40°C for 5-6h. A saturated sodium bicarbonate aqueous solution was added to the system to adjust the pH to weak alkalinity. The excess methanol was removed by vacuum concentration. Ethyl acetate was added to the system for extraction three times. The organic phase was dried over anhydrous sodium sulfate and concentrated. Purification with a silica gel column gave 2.71g of the product. Ms[M+H] ⁺ : 396.2.

3. Synthesis of int2D-4

In a 100 mL flask, int2D-3 (2.71 g, 1.0 eq), N-methylmorpholine (6.9 g, 10 eq) and dichloromethane (110 mL) were added. P-nitrochloroformic acid phenyl ester (2.76 g, 2 eq) was slowly added to the reaction solution at 0°C, and then reacted at room temperature for 2 min. The reaction was complete. Water was added to the reaction solution at 0°C to quench, separate the layers, extract the aqueous phase with ethyl acetate three times, combine the obtained organic phases, dry with anhydrous sodium sulfate and concentrate. Purify with a silica gel column to obtain 3.22 g of the product. Ms[M+H] ⁺ : 561.2.

4. Synthesis of int2D

Add int2D-4 (3.22 g, 1.0 eq) and formic acid (32 mL) to a 100 mL flask. Heat to 80 °C for 1.5 h, cool, concentrate, add water, add saturated sodium bicarbonate aqueous solution to the system and adjust the pH to Weakly alkaline, ethyl acetate was added for extraction three times, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column to obtain 1.2 g of the product. Ms[M+H] ⁺ : 505.1.

Synthesis of intermediate int2E:

1. Synthesis of int2E-2

Compound int2E-1 (500 mg, 2.97 mmol) was dissolved in N,N-dimethylformamide (25 mL), cooled to 0°C, and a solution of sodium hydride (714 mg, 17.86 mmol) in N,N-dimethylformamide was added dropwise. After the addition was completed, the mixture was stirred at 0°C for 15 min, and then a solution of 1,2-dibromoethane (1.68 g, 8.93 mmol) in N,N-dimethylformamide was added dropwise. After the addition was completed, the mixture was slowly warmed to room temperature and stirred for 3 h. The reaction was completed. The mixture was cooled, and an appropriate amount of water was added to quench the reaction. The mixture was then concentrated under reduced pressure and purified by column chromatography to obtain 527 mg of the product, Ms[M+H] ⁺ : 195.0.

2. Synthesis of int2E

Dissolve compound int2E-2 (470 mg, 2.42 mmol) in N,N-dimethylformamide (15 mL), cool to 0°C, add sodium hydride (116 mg, 2.91 mmol) in N,N-dimethylformamide solution dropwise, stir at 0°C for 15 min, then add iodoethane (1.13 g, 7.27 mmol), slowly warm to room temperature, and stir for 2 h. The reaction is complete. Cool, add appropriate amount of water to quench the reaction, then concentrate under reduced pressure, and purify on a thin layer chromatography preparative plate to obtain 233 mg of the product, Ms[M+H] ⁺ : 223.1.

Synthesis of intermediate int2F:

1. Synthesis of int2F-2

Add Raney nickel (2.50 g) to a 250 mL single-mouth bottle and replace water with methanol. Add int2F-1 (5.00 g, 1 eq), methanol (50 mL) and tetrahydrofuran (50 mL) to replace hydrogen. Stir the reaction solution at room temperature for 3 h in a hydrogen atmosphere. The reaction is complete. Filter the reaction solution, wash with methanol, and concentrate the organic phase under reduced pressure. Purify on a silica gel column to obtain 3.9 g of pure product. Ms[M+H] ⁺ : 177.0.

2. Synthesis of int2F-3

Compound int2F-2 (3.90 g, 1.00 eq), cuprous iodide (211 mg, 0.05 eq) and triethylamine (80 mL) were added to a 250 mL single-mouth bottle. Trimethylsilyl acetylene (3.26 g, 1.50 eq) and bis(triphenylphosphine)palladium dichloride (778 mg, 0.05 eq) were added at room temperature to replace nitrogen. The reaction was carried out at 80 ° C for 2 h under a nitrogen atmosphere. The reaction was complete. The reaction solution was cooled to room temperature, filtered, and the filter residue was washed with ethyl acetate. The organic phase was diluted with ethyl acetate, washed with water, and the aqueous phase was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. 4 g of pure product was obtained after purification on a silica gel column. Ms[M+H] ⁺ : 239.1.

3. Synthesis of int2F-4

Add compound int2F-3 (3.50 g, 1.00 eq) and acetonitrile (60 mL) to a 250 mL single-mouth bottle, and add dropwise 1 M potassium tert-butoxide tetrahydrofuran solution (14.71 mL, 1.50 eq) under an ice bath. React at room temperature for 2 h. The reaction is complete. Dilute the reaction solution with a large amount of ethyl acetate, wash with water and separate the layers. Wash with saturated brine, dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure. Purify on a silica gel column to obtain 2 g of pure product. Ms[M+H] ⁺ : 167.0.

4. Synthesis of int2F-5

Dissolve int2F-4 (500 mg, 1.00 eq) in ultra-dry N,N-dimethylformamide (10 mL), cool to 0°C, add sodium hydride (180 mg, 1.50 eq), and react for 30 min in an ice bath. Add iodomethane (515 mg, 1.2 eq) dropwise. React at room temperature for 1 h. The reaction is complete. Quench the reaction with ice water in an ice bath, extract the aqueous phase with ethyl acetate, and separate the layers. Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Obtain 600 mg of crude product. Ms[M+H] ⁺ : 181.0.

5. Synthesis of int2F-6

Add compound int2F-5 (542 mg, 1.00 eq), sodium bicarbonate (759 mg, 3.00 eq) and tert-butyl alcohol (10 mL) to a 50 mL single-mouth bottle and stir at room temperature for 10 min. Slowly add bromine (2.89 g, 6.00 eq) dropwise. React at room temperature for 1 h. The reaction is complete. Add water to quench the reaction, extract the aqueous phase with ethyl acetate, and separate the layers. Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify on a silica gel column to obtain 800 mg of pure product. Ms[M+H] ⁺ : 352.9.

6. Synthesis of int2F-7

Dissolve int2F-6 (750 mg, 1.00 eq) in glacial acetic acid (7 mL), add zinc powder (692 mg, 5.00 eq) at room temperature. React at room temperature for 1 h. The reaction is complete. Add methanol to the reaction solution for dilution, filter and wash the residue with methanol. Concentrate the filtrate under reduced pressure to obtain 500 mg of crude product. Ms[M+H] ⁺ : 197.0.

7. Synthesis of int2F

Dissolve int2F-7 (417 mg, 1.00 eq), diphenyl (vinyl) sulfonium trifluoromethanesulfonate (771 mg, 1.00 eq) and zinc trifluoromethanesulfonate (155 mg, 0.20 eq) in dichloromethane (10 mL), stir at room temperature for 3 min, and slowly drop 1,8-diazabicyclo [5.4.0] undec-7-ene (648 mg, 2.00 eq) into the system. React at room temperature for 10 min. The reaction is complete. Add saturated ammonium chloride aqueous solution to the reaction solution, separate the layers, and extract the aqueous phase with ethyl acetate. Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify on a silica gel column to obtain 350 mg of pure product. Ms [M + H] ⁺ : 223.1.

Synthesis of intermediate int2G:

1. Synthesis of int2G-2

Dissolve int2G-1 (4 g, 1.00 eq) in ultra-dry N,N-dimethylformamide (10 mL), cool to 0°C, add 60% sodium hydride (1.57 g, 1.50 eq), and react for 30 min in an ice bath. Then, add iodomethane (4.47 g, 1.2 eq) dropwise to the reaction system. React at room temperature for 1 h. The reaction is complete. Quench the reaction with ice water in an ice bath, extract the aqueous phase with ethyl acetate, and separate the layers. Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Obtain 5.39 g of crude product. Ms[M+H] ⁺ : 167.0.

2. Synthesis of int2G-3

Add compound int2G-2 (5.39 g, 1.00 eq), sodium bicarbonate (8.15 g, 3.00 eq) and tert-butanol (250 mL) into a 500 mL single-mouth bottle and stir at room temperature for 10 min. Slowly add bromine (31.0 g, 6.00 eq) dropwise. React at room temperature for 1 h. The reaction is complete. Add water to quench the reaction, extract the aqueous phase with ethyl acetate, and separate the layers. Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify on a silica gel column to obtain 2.167 g of pure product. Ms[M+H] ⁺ : 418.7.

3. Synthesis of int2G-4

Dissolve int2G-3 (2.15 g, 1.00 eq) in glacial acetic acid (100 mL), add iron powder (1.44 g, 5.00 eq) at room temperature. React at room temperature for 4 h. The reaction is complete. Add methanol to dilute the reaction solution, filter and wash the residue with methanol. Concentrate the filtrate under reduced pressure and purify by column chromatography to obtain 893 mg of the product. Ms[M+H] ⁺ : 260.9.

4. Synthesis of int2G

Dissolve int2G-4 (300 mg, 1.00 eq), diphenyl (vinyl) sulfonium trifluoromethanesulfonate (414.5 mg, 1.00 eq) and zinc trifluoromethanesulfonate (83.1 mg, 0.20 eq) in dichloromethane (10 mL), stir at room temperature for 3 min, and slowly drop 1,8-diazabicyclo [5.4.0] undec-7-ene (348.1 mg, 2.00 eq) into the system. React at room temperature for 10 min. The reaction is complete. Add saturated ammonium chloride aqueous solution to the reaction solution, separate the layers, and extract the aqueous phase with ethyl acetate. Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify on a silica gel column to obtain 285 mg of the product. Ms [M + H] ⁺ : 286.9.

Synthesis of intermediate int4:

1. Synthesis of int4-2

Dissolve compound int4-1 (10 g, 76.8 mmol) in 80 mL of N,N-dimethylformamide, then add imidazole (15.7 g, 230.4 mmol), cool and stir under ice bath conditions. Slowly add TBDPSCl (23.2 g, 84.5 mmol), then warm the reaction solution to room temperature and stir overnight. The reaction is complete. Add water to the system, extract with ethyl acetate, dry and concentrate the organic phase, and purify by column chromatography to obtain 24.12 g of compound int4-2. Ms[M+H] ⁺ : 369.1

2. Synthesis of compound int4-3

Under nitrogen protection, n-BuLi (52 mL, 2.5 mol/L) and tetrahydrofuran (70 mL) were added to the flask and stirred in a dry ice bath to cool down. Acetonitrile (5.3 g, 128.4 mmol), after the addition is complete, stir for 1 hour. Then add a solution of compound int4-2 (23.62 g, 64.2 mmol) in tetrahydrofuran (47 mL), after the addition is complete, stir at -78°C for 2 hours. The reaction is complete. Add water to the system, extract with ethyl acetate, dry and concentrate the organic phase to obtain 28.66 g of compound int4-3. Use directly in the next step. Ms[M+H] ⁺ : 378.2.

3. Synthesis of compound int4-4

Add tert-butylhydrazine hydrochloride (14.2 g, 114 mmol) and 143 mL of ethanol to a flask, then add triethylamine (11.52 g, 114 mmol) and stir at room temperature for 30 minutes. Add a solution of compound int4-3 (28.66 g, 76 mmol) in ethanol (143 mL), heat to 75°C and react overnight. The reaction is complete. Concentrate the system until no liquid is distilled out, add water and stir, extract with ethyl acetate, dry and concentrate, and purify by column chromatography to obtain 28.77 g of compound int4-4. Ms[M+H] ⁺ : 448.3.

4. Synthesis of compound int4-5

Under nitrogen protection, compound int4-4 (24.27 g, 54.3 mmol) was dissolved in 170 mL of acetonitrile, and stirred in an ice bath to cool. Cbz-cl (18.5 g, 108.6 mmol) was slowly added dropwise, and stirred at room temperature for 1 to 2 hours after the addition was complete. Sodium bicarbonate (14.6 g, 174 mmol) was added, and the temperature was slowly raised to room temperature and stirred overnight. The reaction was complete. The system was filtered, and the filtrate was concentrated until no liquid was distilled out, water was added and stirred, and ethyl acetate was extracted, dried, concentrated, and purified by column chromatography to obtain 24.88 g of compound int4-5. Ms[M+H] ⁺ : 582.3.

5. Synthesis of compound int4-6

Dissolve compound int4-5 (8.28 g, 14.2 mmol) in 82.8 mL of methanol and stir to cool in an ice bath. Slowly add concentrated hydrochloric acid (1.7 mL) dropwise, and stir at room temperature overnight after the addition is complete. The reaction is complete. Add sodium bicarbonate solution to the system to adjust the pH to alkaline, and remove excess methanol under reduced pressure. Extract with ethyl acetate, dry, concentrate, and purify by column chromatography to obtain 3.57 g of compound int4-6. Ms[M+H] ⁺ : 344.2.

6. Synthesis of compound int4-8

Dissolve compound int4-6 (3 g, 8.74 mmol) and compound int4-7 (2.64 g, 13.11 mmol) in 60 mL of dichloromethane. Slowly add pyridine (2.07 g, 26.22 mmol) and 4-dimethylaminopyridine (0.11 g, 0.874 mmol) and react at room temperature for 5 hours. The reaction is complete. Add water to the system and wash the separated liquid. Add saturated sodium chloride solution to the organic phase and wash the separated liquid 5 times. Concentrate the organic phase and purify by column chromatography to obtain 4.16 g of compound int4-8. Ms[M+H] ⁺ : 509.2.

7. Synthesis of compound int4-9

Dissolve compound int4-8 (4.16 g, 8.2 mmol) and isopropylamine (726 mg, 12.3 mmol) in 83 mL of tetrahydrofuran. Add N,N-diisopropylethylamine (3.2 g, 24.6 mmol) and react at room temperature for 5 hours. The reaction is complete. Add water to the system and extract the liquid with ethyl acetate. Add saturated sodium chloride solution to the organic phase and wash the liquid. Concentrate the organic phase and purify by column chromatography to obtain 2.67 g of compound int4-9. Ms[M+H] ⁺ : 429.2.

8. Synthesis of compound int4

Compound int4-9 (2.67 g, 6.2 mmol) was dissolved in 26 mL methanol/26 mL ethyl acetate. Palladium carbon (0.534 g) was added and stirred overnight under hydrogen. The reaction was complete. The filtrate was filtered, concentrated under reduced pressure, and purified by column chromatography to obtain 1.85 g of compound int4. Ms[M+H] ⁺ : 295.2.

Synthesis of intermediate int4A:

1. Synthesis of int4A-2

In a 250mL single-mouth bottle, int4A-1 (4.30g, 1.0eq), (S)-tert-butylsulfenamide (6.06g, 1.0eq), tetraisopropyl titanate (22.8g, 2.0eq) and tetrahydrofuran (86mL) were added respectively, and the mixture was reacted at 70°C for 16 hours under nitrogen protection. After the reaction was completed and cooled to room temperature, the reaction solution was slowly poured into a mixture of saturated sodium bicarbonate and ethyl acetate, extracted with ethyl acetate (50ml x3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography to obtain 5.20g of the product.

2. Synthesis of int4A-3

Add methyl lithium (10 mL, 3.0 eq) and tetrahydrofuran (15 mL) to a 50 mL single-mouth bottle, cool to -70°C, and then dropwise add a solution of int4A-2 (1.00 g, 1.0 eq) in tetrahydrofuran (5 mL). After the addition is complete, keep the temperature at -70°C for 1 hour and then naturally warm to room temperature for 16 hours. After the reaction is complete and the temperature is lowered to 0°C, the reaction solution is slowly poured into a mixture of saturated sodium bicarbonate and ethyl acetate, extracted with ethyl acetate (50 ml x 3), and the organic phases are combined, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography to obtain 272 mg of the product. Ms[M+H] ⁺ : 206.3.

3. Synthesis of int4A

Int4A-3 (272 mg, 1.0 eq) and dichloromethane (2 mL) were added to a 50 mL single-mouth bottle, and after cooling to 0°C, HCl/dioxane (2 mL) was added dropwise. After the addition, the temperature was naturally raised to room temperature and reacted for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure and slurried with ethyl acetate to obtain 147 mg of a crude product. Ms[M+H] ⁺ : 102.1.

Synthesis of intermediate int4B:

1. Synthesis of int4B-2

In a 250mL single-mouth bottle, int4B-1 (4.30g, 1.0eq), (S)-tert-butylsulfenamide (6.06g, 1.0eq), tetraisopropyl titanate (22.8g, 2.0eq) and tetrahydrofuran (86mL) were added respectively, and the mixture was reacted at 70°C for 16 hours under nitrogen protection. After the reaction was completed by TLC monitoring, the reaction solution was slowly poured into a mixture of saturated sodium bicarbonate and ethyl acetate after cooling to room temperature. The organic phases were combined after extraction with ethyl acetate (50ml x3), dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography to obtain 5.32g of the product. MS: 190.3[M+1]

2. Synthesis of int4B-3

Add methyl lithium (20 mL, 3.0 eq) and tetrahydrofuran (30 mL) to a 50 mL single-mouth bottle, cool to -70 ° C, and then drop int4B-2 (2.00 g, 1.0 eq) in tetrahydrofuran (5 mL) solution. After the addition is complete, keep at -70 ° C for 1 hour and then naturally warm to room temperature for 16 hours. After the reaction is completed and cooled to 0 ° C, the reaction solution is slowly poured into a mixture of saturated sodium bicarbonate and ethyl acetate, extracted with ethyl acetate (50 ml x 3), and the organic phases are combined, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography to obtain 1.50 g of the product.

3. Synthesis of int4B

In a 50 mL single-mouth bottle, int4B-3 (1.50 g, 1.0 eq) and dichloromethane (3 mL) were added, and after cooling to 0°C, HCl/dioxane (3 mL) was added dropwise, and the temperature was naturally raised to room temperature for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure and slurried with ethyl acetate to obtain 1.0 g of a crude product. Ms[M+H] ⁺ : 102.1.

Synthesis of intermediate int4C:

1. Synthesis of int4C-2

In a 250 mL single-necked bottle, compound int4C-1 (10 g, 1.00 eq) was dissolved in dichloromethane (100 mL), ethyl acetate (13.05 g, 1.15 eq) was added, and CbzCl (19.14 g, 1.00 eq) was added dropwise under an ice bath. After the addition was completed, the temperature was raised to room temperature and reacted overnight. After the reaction was completed, the mixture was concentrated, water was added, and the mixture was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 14 g of the product, Ms[M+H] ⁺ : 224.1.

2. Synthesis of int4C-3

In a 25 mL single-necked bottle, compound int4C-2 (500 mg, 1.00 eq) was dissolved in tetrahydrofuran (5 mL), and ethyl acetate (906.4 mg, 4.00 eq) was added. The temperature was lowered to -20°C, and MsCl (641.6 mg, 2.5 eq) was added dropwise. The temperature was raised to room temperature and the reaction was allowed to react overnight. After the reaction was completed, water was added to the reaction solution, and the mixture was extracted with ethyl acetate three times. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 742 mg of a crude product, Ms[M+H] ⁺ : 302.1.

3. Synthesis of int4C-4

Compound int4C-3 (742 mg, 1.00 eq), tert-butanol (7 mL), and cesium fluoride (2244.0 g, 6.00 eq) were added to a 25 mL single-necked bottle and reacted at 75 °C overnight. After the reaction was completed, water and ethyl acetate were added and extracted three times. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 225 mg of the product.

4. Synthesis of int4C

In a 25 mL single-necked bottle, compound int4C-4 (225 mg, 1.00 eq) was dissolved in tetrahydrofuran (5 mL), palladium carbon (50 mg) was added, and the mixture was reacted at room temperature for 2 hours under hydrogen. After the reaction was completed, the crude product was directly used for the next step.

Synthesis of intermediate int4D:

1. Synthesis of int4D-2

Dissolve int4D-1 (5 g, 1.00 eq) in tetrahydrofuran (120 mL), add tetraethyl titanate (24.6 g, 1.81 eq) and stir at room temperature for 10 minutes, add (S)-tert-butylsulfenamide (7.2 g, 1.00 eq) and stir at room temperature for 18 h. Concentrate the reaction system in vacuo until no liquid is distilled out, add ethyl acetate, wash the separated liquid with sodium bicarbonate solution, filter and concentrate to obtain 9.5 g, which is directly used for the next step. ¹ H NMR (300 MHz, CDCl ₃ ) δ: ppm 1.15 (s, 9H), 2.15–2.03 (m, 2H), 3.12-3.03 (m, 2H), 3.27-3.11 (m, 1Ｈ), 3.50-3.39 (m, 1H).

2. Synthesis of int4D-4

Dissolve int4D-3 (5 g, 1.00 eq) in chloroform (100 mL), add diethylaminosulfur trifluoride (23 g, 4.0 eq) under nitrogen protection, and stir at 60 °C for 18 h. Add the reaction system dropwise to saturated carbon at 0 °C. The mixture was extracted with dichloromethane for three times in sodium bicarbonate solution and concentrated to obtain 7.26 g of crude product, which was directly used for the next step. ¹ H NMR (300 MHz, CDCl ₃ ) δ: ppm 5.62 (s, 1H), 5.80 (s, 1H), 7.35-7.26 (m, 3H), 7.51-7.43 (m, 2H).

3. Synthesis of int4D-5

Dissolve int4D-4 (5.07 g, 1.00 eq) in dichloromethane (50 mL), add m-chloroperbenzoic acid (20.9 g, 3.4 eq) at 0°C, and stir at room temperature for 18 h. Add the reaction system dropwise into a 0°C sodium thiosulfate solution, extract with dichloromethane for 3 times, concentrate, and purify by column chromatography to obtain 4.95 g of the product, Ms[M+H] ⁺ : 175.0.

4. Synthesis of int4D-6

Dissolve int4D-5 (4.8 g, 1.53 eq) in tetrahydrofuran (30 mL), cool to -78°C under nitrogen protection, slowly drop n-butyl lithium (11 mL, 1.53 eq), stir for 40 minutes after completion. Add a tetrahydrofuran solution of int4D-2 (3.12 g, 1.00 eq) dropwise at -78°C, stir at room temperature for 18 h. Add water dropwise to the reaction system to quench, extract with dichloromethane for 3 times, concentrate, and purify by column chromatography to obtain 2.8 g of the product, Ms[M+H] ⁺ : 348.1.

5. Synthesis of int4D-7

Dissolve int4D-6 (2.5 g, 1.00 eq) in NN-dimethylformamide (450 mL). Add sodium acetate (37.8 g, 64 eq) and acetic acid (57.5 mL) under nitrogen protection and stir at room temperature for 15 minutes. Add magnesium (11.7 g, 67 eq) and react at 65°C for 18 hours. Filter the reaction system, add sodium bicarbonate to the filtrate to adjust the pH to weak alkalinity, extract with ethyl acetate, and separate the liquids. Wash the separated liquids with saturated sodium chloride, concentrate the organic phase in vacuo until no liquid is distilled out, and purify by column chromatography to obtain 0.49 g of the product, Ms[M+H] ⁺ : 208.1.

6. Synthesis of int4D

Dissolve int4D-7 (0.49 g, 1.00 eq) in methanol (5 mL), add 4M hydrogen chloride/1.4-dioxane solution (5 mL), and stir at room temperature for 30 minutes. Concentrate the reaction system in vacuo until no liquid is distilled out, add dichloromethane and methyl tert-butyl ether for slurrying, and filter to obtain 0.22 g of the product, Ms[M+H] ⁺ : 104.1.

Synthesis of intermediate int4E:

1. Synthesis of int4E-2

Dissolve the compound int4E-1 (30 g, 0.322 mol) in 200 mL of ethanol, add potassium hydroxide (72.3 g, 1.289 mol) and 200 mL of water respectively. Heat to 80°C and react for 16 hours. The reaction is complete. Concentrate under reduced pressure, add ethyl acetate to the system, extract and separate, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and purify by column chromatography to obtain 37.4 g of the product, Ms[MH] ⁺ : 111.1.

2. Synthesis of int4E-3

Compound int4E-2 (10 g, 89.2 mmol) was dissolved in 50 mL of dichloromethane, N,N'-carbonyldiimidazole (21.67 g, 133.8 mmol) was added, and the mixture was reacted at room temperature for 10 min, and then dimethylhydroxylamine hydrochloride (13 g, 133.8mmol) and triethylamine (27.02g, 267.5mmol), react at room temperature for 2 hours. The reaction is complete. Water and dichloromethane are added to the system, and the liquid is extracted. The organic phases are combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 13.3g of the product, Ms[M+H] ⁺ : 156.1.

3. Synthesis of int4E-4

Under nitrogen protection, n-butyl lithium (137.1 mL, 2.5 M in hexane) was added to 300 mL of anhydrous tetrahydrofuran, the system was cooled to -65 ° C, and then acetonitrile (15.8 g, 385.6 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred for 1 hour. Then, a solution of compound int4E-3 (13.3 g, 85.7 mmol) in anhydrous tetrahydrofuran (140 mL) was slowly added dropwise, and the mixture was stirred for 2 hours. After the reaction was completed, water was added dropwise at -65 ° C to quench the reaction, and then the mixture was stirred and heated to room temperature. The reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 6.86 g of crude product, which was directly used in the next step. Ms [M + H] ⁺ : 136.1.

4. Synthesis of int4E-5

Tert-butylhydrazine hydrochloride (9.49 g, 76.1 mmol) was dissolved in 70 mL of ethanol, and triethylamine (7.69 g, 76.1 mmol) was added dropwise under ice bath conditions. After the addition was completed, the mixture was heated to room temperature and stirred for 0.5 hours. Then, an ethanol solution (20 mL) of compound int4E-4 (6.86 g, 50.8 mmol) was added, and the reaction system was heated to 75°C and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated, extracted with water and ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 8.5 g of the product, Ms[M+H] ⁺ : 206.2.

5. Synthesis of int4E-6

Compound int4E-5 (8.5 g, 41.4 mmol) was dissolved in 100 mL of acetonitrile, and benzyl chloroformate (14.08 g, 82.8 mmol) was added dropwise under ice-water cooling. After the addition was complete, the mixture was heated to room temperature and stirred for 2 h, and then sodium bicarbonate (11.13 g, 132.48 mmol) was added in batches and stirred for 16 h at room temperature. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with ethyl acetate for several times. The filtrate was concentrated under reduced pressure and then diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 12.8 g of the product, Ms[M+H] ⁺ : 340.2.

6. Synthesis of int4E-7

Under nitrogen protection, compound int4E-6 (1 g, 3 mmol) was dissolved in 10% tetrahydrofuran, and borane dimethyl sulfide solution (1.5 mL) was added dropwise under ice-salt bath conditions. After the addition was complete, sodium hydroxide aqueous solution (5 mL) and 30% hydrogen peroxide solution (1.7 mL) were added successively under ice-salt bath conditions, and the reaction was continued for 2 hours at room temperature. The reaction was quenched with saturated sodium sulfite aqueous solution (5 mL), diluted with water, and extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and dried, and purified by column chromatography to obtain 1.06 g of the product, Ms[M+H] ⁺ : 358.2.

7. Synthesis of int4E-8

Compound int4E-7 (1.06 g, 2.97 mmol) was dissolved in 20 mL of dichloromethane, and p-nitrophenyl chloroformate (896.3 mg, 4.45 mmol), pyridine (702.8 mg, 8.90 mmol), and DMAP (36.2 mg, 0.3 mmol) were added in sequence, and then stirred at room temperature for 16 h. After the reaction was completed, the reaction solution was washed with saturated brine for 4-5 times, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 1.1 g of the product, Ms[M+H] ⁺ : 523.2.

8. Synthesis of int4E-9

Compound int4E-8 (500 mg, 0.96 mmol) was dissolved in 3 mL of tetrahydrofuran, and isopropylamine (84.84 mg, 1.44 mmol) and diisopropylethylamine (370 mg, 2.87 mmol) were added in sequence, and then stirred at room temperature overnight. After the reaction was completed, the reaction system was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with 1 mol/L sodium hydroxide solution, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 273 mg of the product, Ms[M+H] ⁺ : 443.2.

9. Synthesis of int4E

Compound int4E-9 (273 mg, 0.62 mmol) was dissolved in 10 mL methanol, and then wet Pd/C (60 mg, 20 mol%) was added and stirred at room temperature for 2 h under a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain 80 mg of the product, Ms[M+H] ⁺ : 309.2.

Synthesis of intermediate int4F:

1. Synthesis of int4F-2

Compound int4F-1 (5.0 g, 22.42 mmol) was dissolved in N,N-dimethylformamide (20 mL), and N,N'-carbonyldiimidazole (5.4 g, 33.63 mmol) was added. After stirring for 10 minutes, triethylamine (6.8 g, 67.26 mmol) and methoxymethylamine (4.4 g, 44.84 mmol) were added. The mixture was stirred at room temperature for 16 hours. The reaction was complete. An appropriate amount of water was added to quench the reaction, and the organic phase was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. After purification by column chromatography, 6.0 g of the product was obtained, Ms[M+H] ⁺ : 267.1.

2. Synthesis of int4F-3

Compound int4F-2 (6.0 g, 22.56 mmol) was dissolved in tetrahydrofuran (50 mL), nitrogen was replaced, the temperature was lowered to 0°C, methyl Grignard reagent (47.37 mL, 47.37 mmol) was added dropwise, the temperature was slowly raised to room temperature, and stirred for 18 hours. The temperature was lowered, and an appropriate amount of saturated aqueous ammonium chloride solution was added to quench the reaction, the organic phase was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 2.84 g of the product, Ms[M+H] ⁺ : 222.2.

3. Synthesis of int4F-4

Dissolve the compound int4F-3 (0.8 g, 3.62 mmol) in tetrahydrofuran (8 mL), cool to 0°C, add sodium borohydride (150 mg, 3.98 mmol), stir at 0°C for 2 hours after addition, and perform TLC. Cool, add an appropriate amount of saturated aqueous ammonium chloride solution to quench the reaction, extract the organic phase with ethyl acetate, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify by column chromatography to obtain 743 mg of the product, Ms[M+H] ⁺ : 224.2.

4. Synthesis of int4F

Dissolve the compound int4F-4 (743 mg, 3.33 mmol) in methanol solution (15 mL), add palladium carbon (150 mg), stir at room temperature for 1 hour, perform thin layer chromatography, filter through celite, concentrate under reduced pressure, and purify by column chromatography to obtain 160 mg.

Synthesis of intermediate int4G:

1. Synthesis of int4G-2

Dissolve the compound int4G-1 (10 g, 1.0 eq) in 40 mL of NN-dimethylformamide, add N,N'-carbonyldiimidazole (11 g, 1.5 eq) under ice bath and stir for 10 minutes. Add triethylamine (13.6 g, 3.0 eq) and dimethylhydroxylamine hydrochloride (8.8 g, 2.0 eq) and stir at room temperature for 16 hours until the reaction is complete. Add water to the system and extract with ethyl acetate. Wash the organic phase with saturated sodium chloride solution, concentrate in vacuo until no liquid flows out, and purify by column chromatography to obtain 11.78 g of the product. Ms[M+H] ⁺ : 289.1.

2. Synthesis of int4G-3

Dissolve the compound int4G-2 (11.78 g, 1.0 eq) in 236 mL of tetrahydrofuran. Under nitrogen protection, cool and stir. At an internal temperature of -5°C, slowly drop 118 mL of 1 mol/L methylmagnesium bromide solution. After the addition, slowly warm up to room temperature and react for 12 hours. The reaction is complete. Add saturated ammonium chloride solution dropwise under an ice bath to quench the reaction, and extract with ethyl acetate. Wash the organic phase with saturated sodium chloride solution, concentrate in vacuo until no liquid flows out, and purify by column chromatography to obtain 7.4 g of the product. Ms[M+H] ⁺ : 244.1.

3. Synthesis of int4G-4

Dissolve compound int4G-3 (1 g, 1.0 eq) in 20 mL of tetrahydrofuran and add 5 mL of methanol. Stir and cool under ice bath, slowly add sodium borohydride (188 mg, 1.1 eq) and react at 0°C for 1-2 hours. The reaction is complete. Add saturated ammonium chloride solution dropwise under ice bath to quench the reaction, extract with ethyl acetate and separate the liquids. Wash the organic phase with saturated sodium chloride solution, concentrate in vacuo until no liquid flows out, and purify by column chromatography to obtain 1.08 g of the product, Ms[M+H] ⁺ : 224.1.

4. Synthesis of int4G

Dissolve compound int4G-4 (1.08 g, 1.0 eq) in 10 mL of methanol and add 0.01 g of palladium carbon. After hydrogen replacement 3 times, react at room temperature for 1-2 hours. The reaction is complete. Filter the system and vacuum concentrate until no liquid flows out to obtain 340 mg of the product, Ms[M+H] ⁺ : 90.0.

The following intermediates were synthesized using the synthesis method of int1:

Method 1:

Method 2:

Synthesis of intermediate int1E:

1. Synthesis of int1E-1

In a 100mL three-necked flask, tetrahydrofuran (10mL) and int1D-1 (300mg 1eq) were added. The mixture was cooled to 0°C under _N2 protection, and DI ethyl acetate (332mg 4eq) was added. After stirring for 5min, bicyclo[1.1.1]pentane-1-amine hydrochloride (115.48mg 1.5eq) was added and stirred at 40°C for 6h. After the reaction was completed, ethyl acetate (50ml) was added to the reaction solution, and the mixture was washed with 1M sodium hydroxide (30ml x3) and then with saturated sodium chloride (30ml x3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to 260mg.

2. Synthesis of int1E-2

In a 100mL three-necked flask, dichloromethane (10mL), int1E-1 (260mg 1eq) were added, the mixture was cooled to 0°C under _N2 protection, DI ethyl acetate (163.2mg 2eq), (Boc) ₂ O (179.3mg 1.3eq) and DMAP (15.43mg 0.2eq) were added, and the mixture was stirred at room temperature for 2h. After the reaction was completed, saturated sodium chloride (20ml) and ethyl acetate (50ml x3) were added to the reaction solution for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. Pre-TLC purification yielded 220mg.

3. Synthesis of int1E

In a 100 mL single-necked bottle, int1E-2 (220 mg 1 eq), methanol (20 ml), and Pd/C (110 mg) were added. The mixture was stirred at 30 °C for 2 h under a hydrogen atmosphere, then filtered and concentrated to obtain 180 mg of the product.

The following intermediates were synthesized using the synthesis method of int1E:

Specific embodiment:

Example 1

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C1-1

In a 100 mL sealed bottle, int2 (165 mg, 1.0 eq), Pd(OAc) ₂ (15 mg 0.1 eq), XantPhos (60 g, 0.2 eq), Cs ₂ CO ₃ (334 mg, 3 eq), int1 (182 mg, 1.5 eq), and dioxane (8 ml) were added and reacted at 100° C. for 2 h. After the reaction was completed, the product was filtered and dried by spin drying, and purified by preparative chromatography to obtain 200 mg of C1-1.

2. C1 synthesis

C1-1 (200 mg, 1.0 eq) was added to a 50 mL single-mouth bottle, and then formic acid (10 mL) was added at room temperature, and the reaction was carried out at 80°C for 2 h. The reaction was monitored by thin layer chromatography. The pH was adjusted to near neutral with saturated sodium bicarbonate, and then extracted with ethyl acetate (50 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative chromatography to obtain 50 mg of C1. [M+H] ⁺ :425.2, ¹ H NMR (400 MHz, CDCl ₃ )δ:ppm 7.94(d,J＝5.9Hz,1H),7.13(d,J＝5.9Hz,1H),6.39(s,1H),5.87(s,1H),5.21(s,1H),4.58(s,1H),3.81(s,1H),3.65(s,3H),3.20(t,J＝8.2Hz,1H),2 .58–2.46(m,1H),2.15(dt,J＝12.9,6.1Hz,1H),2.00–1.94(m,2H),1.88(s,2H),1.84(s,2H),1.79(t,J＝3.8Hz,2H),1.15(dd,J＝6.6,3.3Hz,6H).

Example 2

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Prepare 50 mg of compound C1 by the following chiral separation method:

Column: Phenomenex 5 μm Cellulose-4 (250 × 30 mm), mobile phase: n-hexane:isopropanol (0.1% TFA) = 60:40, flow rate: 15 mL/min, column temperature: 25° C., to prepare the products: C3 (11 mg, 25.7 min, [M+H] ⁺ : 425.2), C2 (12 mg, 34.4 min, [M+H] ⁺ : 425.2).

Example 3

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C4-1

In a 100mL three-necked flask, add N,N-dimethylformamide (10mL) and int2-5 (200mg 1eq). Cool to 0°C under nitrogen protection, add NaH (60%) (73.2mg 1.5eq), stir for 5min, slowly add deuterated iodomethane (353.6mg 2eq) dropwise, stir at room temperature for 2h. After the reaction is complete, add 40ml of saturated ammonium chloride to the reaction solution, extract with ethyl acetate (30ml x 3), combine the organic phases, dry the organic phases over anhydrous sodium sulfate, filter, and concentrate to obtain 195mg of the product.

¹ H NMR (400MHZ CDCl ₃ ) δ ppm 8.04 (d 5.9, 1H, 6.73 (d 5.9, 1H), 3.93 (, 3H), 1.83 (, 2H), 1.76 (t3.5, 2H).

2. Synthesis of C4-2

In a 100mL three-necked flask, add C4-1 (195mg 1eq) and dichloromethane (10ml), cool to 0℃ under nitrogen protection, add boron tribromide (5ml), stir at 40℃ for 8h and then reduce the pressure. After concentration, dissolve in dichloromethane and add 1ml of water to quench. After concentration, obtain 300mg of crude product and proceed directly to the next step.

3. Synthesis of C4-3

In a 100mL three-necked flask, add C4-2, dichloromethane (10ml), and triethylamine (306mg 3eq). Under nitrogen protection, cool to 0°C, add trifluoromethanesulfonic anhydride (427.38mg 1.5eq)/dichloromethane (2ml), and stir for 5min. After the reaction is completed, add saturated ammonium chloride 40ml to the reaction solution and extract with dichloromethane (30ml x3). After combining the organic phases, dry the organic phases over anhydrous sodium sulfate, filter, concentrate, and separate and purify with a thin layer preparation plate to obtain 100mg of the product.

4. Synthesis of C4-4

C4-3 (100 mg, 1.0 eq), Pd(OAc) ₂ (6.89 mg 0.1 eq), XantPhos (35.57 mg 0.2 eq), Cs ₂ CO ₃ (301 mg 3 eq), int1 (113.72 mg 1.5 eq), and dioxane (8 ml) were added to a 100 mL sealed bottle and reacted at 100° C. for 2 h. After the reaction was completed, the product was filtered, concentrated, and separated and purified by TLC preparation plate to obtain 97 mg of the product.

5. Synthesis of C4

C4-4 (97 mg, 1.0 eq) was added to a 50 mL single-mouth bottle, and then formic acid (10 mL) was added at room temperature and reacted at 80°C for 2 h. After the reaction was completed by TLC monitoring, the pH was adjusted to near neutral with saturated sodium bicarbonate, and then extracted with ethyl acetate (50 ml x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and separated and purified by TLC preparation plate to obtain 32 mg of the product, ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.94(d,J＝5.7,1H),7.09(d,J＝5.7,1H),6.30(s,1H),5.86(s,1H),5.22(s,1H),4.58(s,1H),3.81(s,1H),3.26–3.15(m,1H),2.52(s,1H),2.15(d ,J＝6.4,1H),1.95(s,2H),1.92–1.79(m,4H),1.76(dd,J＝7.5,3.6,2H),1.15(d,J＝6.4,6H).

Prepared by the following chiral separation method: Chromatographic column: Phenomenex 5μm Cellulose-4 (250x 30mm), mobile phase: n-hexane:isopropanol (0.1% TFA) = 60:40, flow rate: 15mL/min, column temperature: 25°C, the products prepared were: C6 (11.6mg, 26.2min), C5 (11.2mg, 35.2min).

Example 4

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C7-1

In a 100 mL sealed bottle, int2 (150 mg, 1.0 eq), Pd(OAc) ₂ (11.27 mg 0.1 eq), XantPhos (53.85 mg 0.2 eq), cesium carbonate (303 mg 2 eq), int3 (179 mg 1.5 eq), and dioxane (8 ml) were added and reacted at 100° C. for 2 h. After the reaction was completed, the product was filtered, concentrated, and separated and purified by TLC preparation to obtain 100 mg of the product.

2. Synthesis of C7 and C8

C7-1 (100 mg, 1.0 eq) was added to a 50 mL single-mouth bottle, and then formic acid (10 mL) was added at room temperature and reacted at 80°C for 2 h. After the reaction was completed by TLC monitoring, the pH was adjusted to near neutral with saturated sodium bicarbonate, and then extracted with ethyl acetate (50 ml x 3) and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and separated and purified by TLC preparation plate to obtain the product C7-2 (40 mg), which was prepared by the following chiral separation method: Chromatographic column: Phenomenex 5μm Cellulose-4 (250x 30mm), mobile phase: n-hexane: isopropanol (0.1% TFA) = 60:40, flow rate: 15mL/min, column temperature: 25°C, the product was prepared:

C8 (12mg, 29.4min), ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.94 (d, J = 5.7, 1H), 7.07 (d, J = 5.7, 1H), 6.46 (s, 1H), 5.85 (s ,1H),5.21(s,1H),5.01(s,1H),4.13(s,1H),3.63(s,3H),3.25–3.14(m,1H),2.50(s,1H),2.31( s,2H),2.14(s,1H),1.96(d,J＝10.1,2H),1.85–1.72(m,6H),1.71–1.59(m,4H).

C7 (11mg, 41.9min), ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.94 (d, J = 5.8, 1H), 7.07 (d, J = 5.3, 1H), 6.40 (s, 1H), 5.85 (s ,1H),5.20(s,1H),5.01(s,1H),4.16(s,1H),3.63(s,3H),3.26–3.13(m,1H),2.50(s,1H),2.31( s,2H),2.14(s,1H),1.94(s,2H),1.86–1.76(m,6H),1.72–1.63(m,4H).

Example 5

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C9-1

In a 25 mL glass tube, int-2 (150 mg, 1 eq), int4 (137 mg, 1 eq), Cs ₂ CO ₃ (456 mg, 3 eq), Xantphos (54 mg, 0.2 eq), Pd(OAc) ₂ (10 mg, 0.1 eq) and 1,4-dioxane (10 mL) were added. The mixture was heated to 100°C in a nitrogen atmosphere and reacted for 2 h. The reaction was monitored and the reaction was complete. The mixture was cooled, filtered, washed with ethyl acetate three times, and the organic phase was concentrated. The mixture was purified by column chromatography and concentrated to obtain 150 mg of the product. Ms[M+H] ⁺ 467.

2. Synthesis of C9

Add C9-1 (150 mg, 1 eq) and formic acid (2 mL) to a 5 mL glass tube. Heat to 80 °C and react for 80 min. Monitor the reaction to completion. Cool and concentrate. Dissolve in ethyl acetate, adjust pH to neutral with saturated sodium bicarbonate aqueous solution, extract three times with ethyl acetate, dry the organic phase over anhydrous sodium sulfate, and concentrate. After TLC preparation, 24 mg of the product was obtained. ¹ HNMR (400MHz, CDCl ₃ ) δppm 7.94 (d, J = 5.6 Hz, 1H), 7.09 (d, J = 5.6 Hz, 1H), 6.34 (s, 1H), 5.89 (s, 1H), 5.04-4.87 (m, 1H), 4.52 (s, 1H), 3.86-3.73 (m, 1H), 3. 64(s,3H),3.15-3.03(m,1H),2.91-2.80(m,2H),2.28-2.16(m,2H),1.86-1.80(m,2H),1.79-1.74(m,2H),1.16(d,J=6.4Hz,6H).Ms[M+H] ⁺ 411.

Example 6

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C10-1

Compound int2 (150 mg, 1.00 eq), int5 (156.6 mg, 1.0 eq), Pd(OAc) ₂ (10.4 mg, 0.1 eq), Xantphos (53.8 mg, 0.2 eq), cesium carbonate (303.5 mg, 2.0 eq) and 1,4-dioxane (3 mL) were added to a 25 mL single-mouth bottle, and the temperature was raised to 100°C under nitrogen protection for 2 hours. The reaction was monitored for completion, filtered, concentrated, and purified by TLC preparation plate to obtain 93 mg of the product. Ms[M+H] ⁺ 509.

2. Synthesis of C10-2

Compound C10-1 (93 mg, 1.00 eq) and formic acid (3 mL) were added to a 25 mL single-mouth bottle, and the temperature was raised to 80°C for reaction for 1 hour. The reaction was monitored for completion, and the mixture was concentrated under reduced pressure. An aqueous solution of sodium bicarbonate was added to the reaction solution to adjust the pH to 7-8, and the mixture was extracted with ethyl acetate three times. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by TLC preparation plate to obtain 31 mg of the product. Ms[M+H] ⁺ : 453.25.

3. Synthesis of C10 and C11

C10-2 (31 mg) was prepared by the following chiral separation method: Chromatographic column: Phenomenex 5μm Amylose-1 (250x 21.2mm), mobile phase: n-hexane:ethanol=7:3, flow rate: 20mL/min, column temperature: 25°C, the product was prepared:

C10 (7.3mg, 17.6min), Ms[M+H] ⁺ : 453.25, ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.94-7.92(d,J＝8Hz,1H),7.10-7.09(d,J＝4Hz,1H),6.40(s,1H),5.86(s,1H),5.21(s,1H),4.46-4.49 (d,J＝12Hz,1H),3.64(s,3H),3.47(s,1H),3.15-3.29(m,1H),2.57-2.49(m,1H),2.12-2.19(m,1H) ,1.94-2.01(m,2H),1.90-1.75(m,6H),1.49-1.58(m,2H),1.32-1.40(m,2H),0.89(m,6H).

C11 (7.5mg, 26.0min), Ms[M+H] ⁺ : 453.25, ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.94-7.92(d,J＝8Hz,1H),7.10-7.09(d,J＝4Hz,1H),6.40(s,1H),5.86(s,1H),5.21(s,1H),4.46-4.49 (d,J＝12Hz,1H),3.64(s,3H),3.47(s,1H),3.15-3.29(m,1H),2.57-2.49(m,1H),2.12-2.19(m,1H) ,1.94-2.01(m,2H),1.90-1.75(m,6H),1.49-1.58(m,2H),1.32-1.40(m,2H),0.89(m,6H).

Example 7

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C12-1

In a 100 mL sealed bottle, int2 (190 mg, 1.0 eq), Pd(OAc) ₂ (22.4 mg 0.1 eq), XantPhos (117.4 mg 0.2 eq), cesium carbonate (975 mg 3 eq), int6 (483 mg 1.5 eq), and 1,4-dioxane (6 mL) were added, and the mixture was reacted at 100°C for 16 h under nitrogen protection. After the reaction was completed, the mixture was cooled to room temperature, filtered, and concentrated to obtain a crude product, which was purified by TLC plate to obtain 210 mg of the product, Ms[M+H] ⁺ : 495.6.

2. Synthesis of C12-2

C12-1 (190 mg, 1.0 eq) was added to a 50 mL single-mouth bottle, and then formic acid (10 mL) was added at room temperature and reacted at 80°C for 2 h. After monitoring the completion of the reaction, the pH value was adjusted to 7-8 with saturated sodium bicarbonate, and then the organic phases were combined after extraction with ethyl acetate (50 ml x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by TLC plate to obtain 80 mg of the product, Ms[M+H] ⁺ : 439.5.

3. Synthesis of C12 and C13

C12-2 (80 mg) was prepared by the following chiral separation method: Chromatographic column: Phenomenex 5μm Cellulose-4 (250X 30mm), n-hexane:isopropanol (0.1% TFA) = 60:40, flow rate: 15mL/min, column temperature: 25°C, the product was prepared:

C12 (30mg, 24.3min), Ms[M+H] ⁺ : 439.5, ¹ H NMR (400MHz, DMSO-d6) δppm 11.98 (s, 1H), 7.84 (s, 1H), 7.82 (d, J = 5.7 Hz,1H),7.24(s,1H),6.88(d,J＝8.4Hz,1H),5.76(s,1H),4.99(s,1H),3.46(s,3H),3.38(dd,J ＝14.6,7.7Hz,1H),3.05(q,J＝8.5Hz,1H),2.48–2. 38(m,1H),2.07–1.98(m,1H),1.88(d,J＝12.5Hz,1H),1.73(d,J＝8.4Hz,2H),1.66–1.57(m,3H),1.54 (q,J＝4.3,3.6Hz,2H),1.39–1.30(m,2H),1.00(d,J＝6.6Hz,3H),0.79(t,J＝7.4Hz,3H).

C13 (25mg, 30.0min), Ms[M+H] ⁺ : 439.5, ¹ H NMR (400MHz, DMSO-d6) δppm 11.97 (s, 1H), 7.84 (s, 1H), 7.82 (d, J = 5.7 Hz,1H),7.24(s,1H),6.88(d,J＝8.3Hz,1H),5.76(d,J＝1.5Hz,1H),4.99(s,1H),3.46(s,3H), 3.39(dt,J＝14.0,6.1Hz,1H),3.14–2.97(m,1H),2.47– 2.40(m,1H),2.04(d,J＝13.9Hz,1H),1.93–1.84(m,1H),1.78–1.68(m,2H),1.67–1.57(m,3H),1.54(q, J＝4.2, 3.6Hz, 2H), 1.34 (d, J＝7.4Hz, 2H), 0.99 (d, J＝6.6Hz, 3H), 0.81 (t, J＝7.4Hz, 3H).

The following compounds were synthesized using the above synthesis method:

Example 8

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C31-1

Compound int15 (150 mg, 1.00 eq), int2B (231 mg, 1.0 eq), palladium acetate (16 mg, 0.1 eq), XantpHos (83 mg, 0.2 eq), cesium carbonate (715 mg, 3.0 eq) and 1,4-dioxane (23 mL) were added to a 25 mL single-necked bottle. The temperature was raised to 100 °C under nitrogen protection and the reaction was carried out for 2 hours. After the reaction was completed, the mixture was filtered, concentrated and purified by TLC preparative plate to obtain 230 mg of the product, Ms[M+H] ⁺ : 493.2.

2. Synthesis of C31-2

Compound C31-1 (230 mg, 1.00 eq) and formic acid (23 mL) were added to a 25 mL single-mouth bottle, and the temperature was raised to 80°C for reaction for 1 hour. After the reaction was completed, the mixture was concentrated under reduced pressure. An aqueous solution of sodium bicarbonate was added to the reaction solution to adjust the pH to 7-8. The mixture was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by TLC preparative plate to obtain 97 mg of the product, Ms[M+H] ⁺ : 437.2.

3. Preparation of C31 and C32

C31-2 (97 mg) was separated by chiral high performance liquid chromatography to obtain the product.

The separation conditions are as follows:

Instrument: Agilent 1260 InfinityⅡ

Column: PHenomenex 5μm Amylose-1(250X 21.2nm)

Mobile phase: n-hexane:ethanol=6:4

Flow rate: 20mL/min, detection wavelength: 254nm

Column temperature: 25°C, solvent: isopropanol

C31 (14.4min, 13.95mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm7.92 (d, J = 5.6Hz, 1H), 7.08 (d, J = 5.6Hz, 1H), 6.39 (s, 1H), 5.85(s,1H),5.15(d,J＝37.2Hz,2H),3.63(s,3H),3.19(s,1H),2.52(dt,J＝15.2,8.0Hz,1H),2.14(s ,1H),1 .94(s,2H),1.84(q,J＝3.6,2.8Hz,3H),1.76(q,J＝4.8,3.6Hz,2H),1.35(s,3H),1.25(tt,J＝4.8 ,2.0Hz,1H),0.91-0.80(m,1H),0.74(d,J=4.4Hz,2H),0.62(s,2H).Ms[M+H] ⁺ : 437.2.

C32 (20.6min, 15.76mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.92 (d, J = 5.6Hz, 1H), 7.08 (d, J = 5.6Hz, 1H), 6.40 (s, 1H), 5.85 (s,1H),5.36-5.01(m,2H),3.63(s,3H),3.19(s,1H),2.52(dt,J＝14.8,8.0Hz,1H),2.14(s,1H), 1 .94(s,2H),1.87-1.80(m,3H),1.78-1.72(m,2H),1.35(s,3H),1.26(ddd,J＝7.2,5.6,2.0Hz,1H),0.88 -0.82(m,1H),0.74(t,J=3.2Hz,2H),0.61(s,2H).Ms[M+H] ⁺ : 437.2.

Example 9

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C35-1

In a 25 mL reaction bottle, int2B (150 mg, 1.0 eq), int16 (300 mg, 1.3 eq), palladium acetate (17 mg, 0.1 eq), XantPHos (83 mg, 0.2 eq), carbonate (700 mg, 3.0 eq), 1,4-dioxane (15 mL) were added and reacted at 100°C for 3 h. After the reaction was completed, water was added for dilution, and ethyl acetate was extracted three times. The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified to obtain 200 mg of the product.

2. Synthesis of C35-2

C35-1 (200 mg, 1.0 eq) was added to a 25 mL reaction bottle, and then formic acid (5 mL) was added, and the reaction was carried out at 80°C for 2 h. After the reaction was completed, a saturated sodium bicarbonate aqueous solution was slowly added to adjust the pH to 8-9, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified to obtain 55 mg of the product.

3. Synthesis of C35 and C36

C35-2 (55 mg) was chiral separated according to the following separation conditions:

Instrument: Agilent 1260 InfinityⅡ

Column: PHenomenex 5μm Amylose-1(250X 21.2nm)

Mobile phase: n-hexane:ethanol=6:4

Flow rate: 15mL/min, detection wavelength: 254nm

Column temperature: 25°C, solvent: isopropanol

C36 (27.1min, 18mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm7.92 (d, J = 5.6Hz, 1H), 7.25 (d, J = 5.6Hz, 1H), 6.72-6.55 (m, 1H) ,5.85(s,1H),5.87-5.12(m,2H),4.91-4.77(m,1H),3.66(s,3H),3.31-3.21(m,1H),2.81-2.48(m,3H) ,2.26-1.73(m,8H),0.92-0.73(m,4H).Ms[M+H] ⁺ : 423.4.

C35 (37.2min, 19mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm7.92 (d, J = 5.6Hz, 1H), 7.25 (d, J = 5.6Hz, 1H), 6.89-6.71 (m, 1H) ,5.85(s,1H),5.12-5.87(m,2H),4.91-4.77(m,1H),3.65(s,3H),3.31-3.21(m,1H),2.81-2.48(m,3H) ,2.28-1.78(m,8H).Ms[M+H] ⁺ : 423.4.

Example 10

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C63-1

Add int2F (150 mg, 1.00 eq), int7 (225 mg, 1.00 eq), cesium carbonate (661 mg, 3.00 eq), XantpHos (78 mg, 0.20 eq), palladium acetate (15 mg, 0.10 eq) and 1,4-dioxane (5 mL) to a sealed glass tube and replace the nitrogen. Heat to 100°C and react for 2 h. The reaction is complete. Cool the reaction solution, filter, wash the solid with ethyl acetate, and concentrate the filtrate under reduced pressure. Purify the product on a TLC preparative plate to obtain 250 mg of pure product. Ms[M+H] ⁺ : 519.3.

2. Synthesis of C63

Compound C63-1 (200 mg, 1.00 eq) was dissolved in formic acid (6 mL), heated to 80 °C and stirred for 1.5 hours. The reaction was complete. The reaction solution was cooled to room temperature and concentrated under reduced pressure. It was redissolved in ethyl acetate, the pH was adjusted to alkaline with a saturated aqueous solution of sodium bicarbonate, and the aqueous phase was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The product was purified by TLC preparation plate to give 57 mg. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 6.93 (s, 1H), 6.23 (s, 1H), 5.83 (s, 1H), 5.33-5.14 (m, 2H), 3.59 (s, 3H), 3.26-3.14 (m, 1H), 2.58-2.47 (m, 1H), 2.41 (s, 1H), 2.40 (s, 3H), 2.20-2.09 (m, 1H), 2.07-1.91 (m, 7H), 1.90-1.76 (m, 4H), 1.75-1.69 (m, 2H), 1.68-1.62 (m, 1H). Ms [M+H] ⁺ : 463.2.

The following compounds were synthesized using the above synthesis method:

Embodiment 11

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C37-1

In a 50 mL flask, int2C (280 mg, 1.0 eq), tetrahydrofuran (10 mL), (S)-1-cyclopropylethylamine (95 mg, 2.0 eq) and N,N-diisopropylethylamine (285 mg, 4.0 eq) were added and reacted at 50°C for 2 h. After the reaction, water was added to the reaction solution, and then extracted with ethyl acetate three times, the organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified to obtain 50 mg of the product.

2. Synthesis of C37 and C38

C37-1 (50 mg) was prepared by chiral separation using the following method:

Instrument: Agilent 1260 InfinityⅡ

Column: PHenomenex 5μm Amylose-1(250X 21.2nm)

Mobile phase: n-hexane:ethanol=6:4

Flow rate: 15mL/min, detection wavelength: 254nm

Column temperature: 25°C, solvent: isopropanol

After chiral separation, the following was obtained:

C38 (21.1min, 16mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.95 (d, J = 5.6Hz, 1H), 7.09 (d, J = 5.6Hz, 1H), 6.45 (br, 1H), 5.86 ( s,1H),5.23(m,1H),4.86(m,1H),3.66(s,3H),3.12(q,J＝8.0Hz,1H),3.10-3.05(m,1H),2.59-2.30 (m,2H),2.14-1.58(m, 8H),0.95-0.79(m,4H),0.52-0.23(m,4H).Ms[M+H] ⁺ : 451.7.

C37 (28.9min, 17mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.94 (d, J = 5.6Hz, 1H), 7.18 (d, J = 5.6Hz, 1H), 6.45 (br, 1H), 5.86 ( s,1H),5.22(m,1H),4.86(m,1H),3.66(s,3H),3.12(q,J＝8.0Hz,1H),3.10-3.05(m,1H),2.59-2.30 (m,2H),2.14-1.71(m,8H),0.94-0.78(m,4H),0.53-0.21(m,4H).Ms[M+H] ⁺ : 451.7.

Example 12

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C61

Add compound int2D (80 mg, 1.00 eq), (S)-(+)-1-methoxy-2-propylamine (21.2 mg, 1.5 eq) and tetrahydrofuran (1.6 mL) to a 25 mL single-mouth bottle, and then add N,N-diisopropylethylamine (61 mg, 3.00 eq). Heat to 50°C and react for 1 to 2 hours until the reaction is complete. The system is dried and water is added, extracted with ethyl acetate, separated, and the organic phase is concentrated. The product is separated by high performance liquid chromatography to obtain 26.75 mg, Ms[M+H] ⁺ : 455.3. ¹ H NMR (400 MHz, CDCl ₃ )δppm7.93(d,J＝5.6Hz,1H),7.09(d,J＝5.6Hz,1H),6.38(s,1H),5.86(s,1H),5.19(dt,J＝6.4,3.9Hz,1H),4.94(s,1H),3.87(s,1H),3.64(s,3H),3.35 (s,5H),3.20(s,1H),2.51(ddd,J＝15.1,9.3,6.4Hz,1H),2.15(d,J＝6.0Hz,1H),1.94(d,J＝6.0Hz,2H),1.91–1.79(m,4H),1.79–1.63(m,3H),1.17(d,J＝ 6.8Hz,3H).

Example 13

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of compound C101-1

Dissolve the compounds int2D (150 mg, 1.0 eq) and int4G (40 mg, 1.5 eq) in tetrahydrofuran (15 mL), then add N,N-diisopropylethylamine (115 mg, 3.00 eq). Heat to 50°C and react for 20 minutes. The system is spin-dried and purified by TLC preparation plate to obtain 100 mg of the product, Ms[M+H] ⁺ : 455.3

2. Synthesis of C101 and C102

Compound C101-1 was prepared and resolved:

The separation conditions are as follows:

Instrument: Agilent 1260 InfinityⅡ

Column: PHenomenex 5μm Amylose-1(250X 21.2nm)

Mobile phase: n-hexane: ethanol = 60:40

Flow rate: 20mL/min, detection wavelength: 254nm

Column temperature: 25°C, solvent: isopropanol

C101 (35.2min, 42.34mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.88 (d, J = 5.6Hz, 1H), 7.02 (d, J = 5.6Hz, 1H), 6.65 (s, 1H), 5.83 (s,1H),5.34–5.10(m,2H),3.87(s,1H),3.80–3.65(m,1H),3.59(s,3H),3.25–3.09(m,1H),2.56–2.36 (m,1H),2.12(s,1H),2.03–1.67(m,9H),1.29–0.96(m,7H).Ms[M+H] ⁺ ：455.3

C102 (44.8min, 16.54mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.91 (d, J = 5.6Hz, 1H), 7.04 (d, J = 5.6Hz, 1H), 6.56 (s, 1H), 5.84 (s,1H),5.17(d,J＝19.2Hz,2H),3.61(s,5H),3.20(s,1H),2.51(s,2H),2.14(s,1H),2.02–1.67( m,8H),1.17(dd,J＝16.0,4.8Hz,7H).Ms[M+H] ⁺ : 455.3

Embodiment 14

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C117

Compound int4C (5 mL, 1.5 eq) was added with int2D (75 mg, 1.0 eq) and N,N-diisopropylethylamine (38.4 mg, 2.0 eq) and reacted at 50°C for 2 hours. Water was added to the system, extracted with ethyl acetate three times, washed with 1M sodium hydroxide aqueous solution three times, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by TLC preparative plate to obtain 35 mg of the product, Ms[M+H] ⁺ : 457.2, ¹ H NMR (400MHz, DMSO-d6) δ12.00(s,1H),7.93–7.75(m,2H),7.26-7.27(d,J＝4Hz,1H),7.04-7.01(d,J＝12Hz,1H),5.77(s,1H),5.01-4.99(d,J＝7.1Hz,1H),4. 50-4.47(t , 1.08-1.06(m,3H).

The following compounds were synthesized using the above synthesis method:

Embodiment 15

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C14-1

In a 100mL three-necked flask, N,N-dimethylformamide (10mL) and int2A-4 (150mg 1eq) were added. The mixture was cooled to 0°C under _N2 protection, and cesium carbonate (467g 2eq) was added. After stirring for 5min, deuterated iodomethane (156mg 1.3eq) was slowly added dropwise. The mixture was stirred at room temperature for 0.5h. After the reaction was completed, saturated ammonium chloride 40ml was added to the reaction solution, and the mixture was extracted with ethyl acetate (30ml x3). After the organic phases were combined, the organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column to obtain 35mg of the product.

2. Synthesis of C14

C14-1 (62 mg 1 eq), int1E (35 mg 1 eq) and DMSO (5 ml) were added to a 100 mL single-mouth bottle. After stirring and dissolving, potassium tert-butoxide (55.47 mg 3 eq) was added and stirred at room temperature for 20 min. After the reaction was completed, saturated sodium chloride (20 ml) was added to the reaction solution, and ethyl acetate (50 ml x 3) was used for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 9.5 mg of the product, Ms[M+H] ⁺ : 453.2, ¹ H NMR(400MHz,DMSO-d6)δ12.11(s,1H),8.53(s,1H),8.30(s,1H),7.78(s,1H),6.27(s,1H),5.01(s,1H),3.09(s,1H),2.35(s,1H),2.04(s,1H),1.91(m, 8H),1.75(m,4H),1.65(s,3H).

Example 16

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C33-1

Compound int2E (130 mg, 0.57 mmol), int1 (176 mg, 0.57 mmol), palladium acetate (13 mg, 0.06mmol), XantpHos (66mg, 0.11mmol), cesium carbonate (560mg, 1.72mmol) and 1,4-dioxane (15mL) were added and the temperature was raised to 100°C under nitrogen protection for 2 hours. After the reaction was completed, the mixture was filtered, concentrated and purified by TLC preparation plate to obtain 325mg of the product, Ms[M+H] ⁺ : 495.3.

2. Synthesis of C33-2

Compound C33-1 (180 mg, 0.36 mmol) was dissolved in formic acid solution (10 mL), heated to 80°C and stirred for 1.5 h. After the reaction was completed, the mixture was concentrated under reduced pressure. An aqueous solution of sodium bicarbonate was added to the reaction solution to adjust the pH to 7-8. The mixture was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by TLC preparative plate to obtain 55 mg of the product, Ms[M+H] ⁺ : 439.2.

3. Preparation of C33 and C34

C33-2 (55 mg) was prepared by chiral HPLC separation

The separation conditions are as follows:

Instrument: Agilent 1260 InfinityⅡ

Column: PHenomenex 5μm Cellulose-4 (250X 21.2nm)

Mobile phase: n-hexane: isopropanol (0.1% TFA) = 7:3

Flow rate: 20mL/min, detection wavelength: 254nm

Column temperature: 25°C, solvent: isopropanol

C33 (17.9min, 14.8mg) ¹ H NMR (400MHz, DMSO-d6) δppm 11.97 (s, 1H), 7.84 (d, J = 5.6Hz, 1H), 7.81 (s, 1H), 7.22 (d, J ＝5.6Hz,1H),6.94(d,J＝5.6Hz,1H),5.75-5.73(m,1H),5.02-4.96(m,1H),4.04(q,J＝7.2Hz,2H),3.62 -3.52(m,1H),3.10-3.01(m ,1H),2.48-2.40(m,1H),2.07-1.98(m,1H),1.95-1.85(m,1H),1.76-1.69(m,2H),1.65-1.62(m,2H),1.62 -1.57(m,1H),1.57-1.53(m,2H),1.09(t,J＝7.2Hz,3H),1.04-0.99(m,6H).Ms[M+H] ⁺ : 439.2.

C34 (28.4min, 10.4mg) ¹ H NMR (400MHz, DMSO-d6) δppm 11.97 (s, 1H), 7.84 (d, J = 5.6Hz, 1H), 7.81 (s, 1H), 7.22 (d, J ＝5.6Hz,1H),6.94(d,J＝5.6Hz,1H),5.75-5.73(m,1H),5.03-4.94(m,1H),4.04(q,J＝7.2Hz,2H),3.63 -3.51(m,1H),3.11-3.00(m ,1H),2.48-2.38(m,1H),2.07-1.97(m,1H),1.95-1.83(m,1H),1.77-1.69(m,2H),1.65-1.62(m,2H),1.62 -1.57(m,1H),1.57-1.53(m,2H),1.09(t,J＝7.2Hz,3H),1.04-0.99(m,6H).Ms[M+H] ⁺ : 439.2.

Embodiment 17

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C43-1

Compound int2B (126.3 mg, 1.00 eq), int4E (187 mg, 1.0 eq), palladium acetate (13.6 mg, 0.1 eq), XantpHos (70.2 mg, 0.2 eq), cesium carbonate (395.9 mg, 2.0 eq) and 1,4-dioxane (5 mL) were added to a 25 mL single-mouth bottle, and the temperature was raised to 100° C. under nitrogen protection for 2 hours. After the reaction was completed, the mixture was filtered, concentrated, and purified by TLC preparative plate to obtain 240 mg of the product. Ms[M+H] ⁺ : 481.3.

2. Synthesis of C43-2

Compound C43-1 (240 mg, 1.00 eq) and formic acid (6 mL) were added to a 25 mL single-mouth bottle, and the temperature was raised to 100°C for reaction for 30 minutes. After the reaction was completed, the mixture was concentrated under reduced pressure, and an aqueous solution of sodium bicarbonate was added to the reaction solution to adjust the pH to 7-8. The mixture was extracted with ethyl acetate three times, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by TLC preparative plate to obtain 80 mg of the product. Ms[M+H] ⁺ : 425.2.

3. Synthesis of C43 and C44

C43-2 (80 mg) was prepared by chiral high performance liquid chromatography separation under the following conditions:

Instrument: Agilent 1260 InfinityⅡ

Column: PHenomenex 5μm Amylose-1(250X 21.2nm)

Mobile phase: n-hexane:ethanol=2:8

Flow rate: 15mL/min, detection wavelength: 254nm

Column temperature: 25°C, solvent: isopropanol

C44 (14.0min, 32mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.91-7.89(d,J＝8Hz,1H),7.07-7.06(d,J＝4Hz,1H),6.67(s,1H),5.84(s,1H),4.60(s,1H),4.09-4.07 (d,J＝8Hz,2H),3.82-3.80(m,1H),3.63(s,3H),3.47-3.38(m,1H),2.63(m,1H),2.53-2.46(m,J＝ 2H),2.05-1.98(m,2H),1.84-1.79(m,2H),1.77-1.73(m,2H),1.25(s,1H),1.17-1.15(d,J＝8Hz,6H). Ms[M+H] ⁺ :425.2.

C43 (22.9min, 22mg) ¹ H NMR (400MHz, CDCl ₃ ) δppm 7.94-7.92(d,J＝8Hz,1H),7.11-7.10(d,J＝4Hz,1H),6.43(s,1H),5.92(s,1H),4.55(s,1H),4.17-4.15 (d,J＝8Hz,2H),3.83-3.81(m,1H),3.64(s,1H),3.60-3.51(m,1H),2.72(m,1H)2.30-2.26(m,4H), 1.85-1.81(m,2H),1.79-1.74(m,2H),1.25(m,1H),1.17-1.16(d,J=8Hz,6H).Ms[M+H] ⁺ : 425.2.

Embodiment 18

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C70-2

Compound C70-1 (3.00 g, 29.7 mmol) was dissolved in N,N-dimethylformamide (50 mL), cooled to 0°C, potassium carbonate (8.2 g, 59.4 mmol) and benzyl bromide (6.1 g, 35.6 mmol) were added, and the temperature was slowly raised to room temperature and stirred for 4 hours. The reaction was complete. The temperature was lowered, and an appropriate amount of water was added to quench the reaction. The product was extracted with ethyl acetate three times, and the ethyl acetate phase was washed with saturated brine, then concentrated under reduced pressure, and purified by column chromatography to obtain 3.3 g of the product, Ms[M+H] ⁺ : 192.0.

2. Synthesis of C70-3

Compound C70-2 (2.0 g, 10.5 mmol) was dissolved in acetonitrile (23 mL), and N-bromosuccinimide (2.1 g, 11.5 mmol) was added. After the addition, the mixture was stirred at room temperature for 3 days. The reaction was complete. An appropriate amount of water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 1.93 g of the product, Ms[M+H] ⁺ : 270.0.

3. Synthesis of C70-4

Compound C70-3 (1.93 g, 7.17 mmol), cyclopropylboronic acid (6.17 g, 71.75 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (525 mg, 0.72 mmol), and cesium carbonate (4.68 g, 14.35 mmol) were dissolved in 1,4-dioxane/water (v: 4:1, 12 mL). The temperature was raised to 100°C under nitrogen protection and the reaction was carried out for 12 hours. After the reaction was completed, the mixture was filtered, concentrated, and purified by TLC preparative plate to obtain 1.04 g of the product, Ms[M+H] ⁺ : 232.1.

4. Synthesis of C70-5

Compound C70-4 (1.04 g, 4.50 mmol) was dissolved in concentrated hydrochloric acid (12 mL), and the temperature was raised to 50°C for reaction for 5 hours. The reaction was complete. The mixture was concentrated under reduced pressure and subjected to reverse phase column chromatography to obtain 537 mg of the product, Ms[M+H] ⁺ : 142.0.

5. Synthesis of C70-6

Compound C70-5 (0.5 g, 3.55 mmol), int1-7 (1.26 g, 3.55 mmol), triphenylphosphine (1.39 g, 5.32 mmol) were dissolved in tetrahydrofuran (50 mL), cooled to 0°C, and a solution of diisopropyl azodicarboxylate (1.07 g, 5.32 mmol) in tetrahydrofuran (10 mL) was added dropwise. After addition, the mixture was stirred at 0°C for 5 minutes, and then heated to 50°C and stirred for 2 hours. The reaction was complete. An appropriate amount of water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. Column chromatography was used for purification to obtain 2.30 g of a crude product, Ms[M+H] ⁺ : 481.2.

6. Synthesis of C70-7

The crude compound C70-6 (1.00 g) was dissolved in 1,4-dioxane/water (v: 10:1, 10 mL), and lithium hydroxide (50 mg, 2.08 mmol) was added, and the temperature was raised to 80°C and stirred for 5 hours. The reaction was complete. The mixture was concentrated under reduced pressure and purified by column chromatography to obtain 471 mg of the product, Ms[M+H] ⁺ : 347.2

7. Synthesis of C70-8

Compound C70-7 (400 mg, 1.00 eq), int2B (240 mg, 1.00 eq), palladium acetate (103 mg, 0.40 eq), XantpHos (534 mg, 0.80 eq), cesium carbonate (1.13 g, 3.00 eq) and 1,4-dioxane (16 mL) were added to a 25 mL single-necked bottle. The temperature was raised to 100 °C under nitrogen protection and the reaction was carried out for 2 hours. After the reaction was completed, the mixture was filtered, concentrated and purified by TLC preparative plate to obtain 200 mg of the product, Ms[M+H] ⁺ : 519.3.

8. Synthesis of C70-9

Compound C70-8 (200 mg, 1.00 eq) and trifluoroacetic acid/water (10 mL, v: 20:1) were added to a 25 mL single-mouth bottle, and the temperature was raised to 70°C for reaction for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure. An aqueous solution of sodium bicarbonate was added to the reaction solution to adjust the pH to 7-8. The mixture was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by TLC preparative plate to obtain 150 mg of the product, Ms[M+H] ⁺ : 463.2.

9. Synthesis of C70 and C71

C70-9 (150 mg) was prepared by chiral high performance liquid chromatography separation under the following conditions:

Instrument: Agilent 1260 InfinityⅡ

Column: PHenomenex 5μm Amylose-1(250X 21.2nm)

Mobile phase: n-hexane: isopropanol (0.1% TFA) = 7:3

Flow rate: 20mL/min, detection wavelength: 254nm

Column temperature: 25°C, solvent: isopropanol

C70 (26.3min, 24.3mg) ¹ H NMR (400MHz, DMSO-d6) δ12.02 (s, 1H), 8.27 (s, 1H), 7.84 (s, 1H), 7.81 (d, J = 5.6Hz, 1H),7.24(d,J＝5.6Hz,1H),5.78(s,1H),5.36-5.30(m,1H),3.45(s,3H),3.20-3.12(m,1H),2. 65-2.57(m,1H),2.12-2.01(m,2H),1.97-1.90(m,1H),1.88-1.78(m,2H),1.73-1.67(m,1H),1.66-1.60(m ,2H),1.57-1.50(m,2H),0.83-0.78(m,2H),0.65-0.61(m,2H).Ms[M+H] ⁺ : 463.2.

C71 (33.3min, 25.1mg) ¹ H NMR (400MHz, DMSO-d6) δ12.02 (s, 1H), 8.27 (s, 1H), 7.85 (s, 1H), 7.81 (d, J = 5.6Hz, 1H),7.24(d,J=5.6Hz,1H),5.78(s,1H),5.35-5.30(m,1H),3.46(s,3H),3.21-3.12(m,1H),2. 66-2.58(m,1H),2.11-2.01(m,2H),1.97-1.90(m,1H),1.87-1.79(m,2H),1.74-1.67(m,1H),1.65-1.60(m ,2H),1.56-1.51(m,2H),0.83-0.78(m,2H),0.65-0.61(m,2H).Ms[M+H] ⁺ : 463.2.

Embodiment 19

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C85-2

Compound C85-1 (3.0 g, 14.22 mmol) was dissolved in N, N-dimethylformamide (50 mL), cooled to 0°C, and sodium hydride (3.41 g, 85.31 mmol) was added in batches. After addition, the mixture was stirred at 0°C for 15 minutes, and then a solution of 1, 2-dibromoethane (7.9 g, 42.65 mmol) in N, N-dimethylformamide was added dropwise. After addition, the mixture was slowly warmed to room temperature and stirred for 2 hours. The reaction was complete. The mixture was cooled, and an appropriate amount of water was added to quench the reaction. The mixture was then concentrated under reduced pressure and purified by column chromatography to obtain 948 mg of the product, Ms[M+H] ⁺ : 238.0.

2. Synthesis of C85-3

Compound C85-2 (940 mg, 3.95 mmol) was dissolved in N,N-dimethylformamide (40 mL), cooled to 0°C, and sodium hydride (190 mg, 4.74 mmol) was added in batches. After addition, the mixture was stirred at 0°C for 30 minutes, and then iodomethane (1.7 g, 11.85 mmol) was added dropwise. After addition, the temperature was slowly raised to room temperature and stirred for 2 hours. The reaction was complete. The mixture was cooled, and an appropriate amount of water was added to quench the reaction, and then the mixture was concentrated under reduced pressure. The product was purified by TLC preparation plate to obtain 968 mg, Ms[M+H] ⁺ : 252.0.

3. Synthesis of C85-4

Compound C85-3 (150 mg, 0.60 mmol), int7 (158 mg, 0.49 mmol), palladium acetate (13 mg, 0.06 mmol), XantpHos (69 mg, 0.12 mmol), cesium carbonate (584 mg, 1.79 mmol) and 1,4-dioxane (15 mL) were heated to 100°C under nitrogen protection for 2 hours. After the reaction was completed, the mixture was filtered, concentrated and purified by TLC preparation plate to obtain 264 mg of the product, Ms[M+H] ⁺ : 504.3.

4. Synthesis of C85

Compound C85-4 (219 mg, 0.44 mmol) was dissolved in acetic acid/water solution (v:20:1) (15 mL), heated to 110°C and stirred for 2 days. After the reaction was completed, it was concentrated under reduced pressure. An aqueous solution of sodium bicarbonate was added to the reaction solution to adjust the pH to 7-8. The solution was extracted with ethyl acetate three times. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by TLC preparative plate to obtain 20 mg of the product. ¹ H NMR(400MHz,DMSO-d6)δppm 11.52(s,1H),7.74(s,1H),7.32(s,1H),7.07(d,J＝8.0Hz,1H),6.88(t,J＝8.0Hz,1H),6.68(d,J＝7.2Hz,1H),5.39(s,1H),4.96(s,1H),3.35(s,3H),3.09-2.90(m,1H),2.47-2.37(m,1H),2.37-2.26m,1H),2.03-1.93(m,1H),1.92-1.80(m,7H),1.73-1.61(m,2H),1.59-1.45(m,5H).Ms[M+H] ⁺ :448.2.

Embodiment 20

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C89-2

In a 250mL three-necked flask, add water (10mL), methylamine hydrochloride (9.2mg 2eq), and sodium bicarbonate (11.4g 5eq). After stirring for 1h under _N2 protection, add C89-1 (5g 1.3eq) and butyl acetate (100ml). Stir at 60℃ for 15h. After the reaction is completed, add 40ml of saturated ammonium chloride to the reaction solution, extract with ethyl acetate (30ml x3), combine the organic phases, dry the organic phases over anhydrous sodium sulfate, filter, concentrate, and purify by column to obtain 1.8g of the product.

2. Synthesis of C89-3

Dissolve dimethyl malonate (7.6g 5eq) in N,N-dimethylformamide (34ml), heat to 40℃, add sodium hydride (1.9g 5eq), stir and react for 0.5h, then add C89-2 (1.7g 1eq) and slowly heat to 100℃, keep warm and react for 16h. After the reaction, cool to room temperature, remove N,N-dimethylformamide by vacuum distillation, add water (20mL), extract and combine organic phases with ethyl acetate (100ml x3), wash with water (25ml x2), combine aqueous phases and adjust pH to 5-6 with concentrated hydrochloric acid, extract and combine organic phases with ethyl acetate (100ml x3), spin dry to obtain 1g of product, ^1H NMR (400MHz, DMSO) δ＝10.98(s,1H),8.03(s,1H),4.15(d,J＝7.1,2H),1.18(s,3H).

3. Synthesis of C89-4

Add C89-3 (1 g) acetic acid (10 mL) into a 50 mL reactor, add concentrated hydrochloric acid (10 mL) under stirring, then slowly raise the temperature to 100 ° C, keep stirring and react for 15 hours, then cool the reaction solution to room temperature, spin dry to obtain 800 mg of crude product, which is directly used in the next step reaction.

4. Synthesis of C89-5

C89-4 (800 mg, 1.0 eq), DMSO (20 ml), DBU (3.3 g 4 eq), Zn (OTf) ₂ (0.84 g 0.4 eq) were added to a 100 mL single-mouth bottle, stirred at 25 ° C for 5 min, and diphenyl (vinyl) sulfonium trifluoromethanesulfonate (1.98 g 1 eq) was added. The reaction was carried out at 25 ° C for 10 min. After the reaction was completed, saturated sodium chloride solution (60 ml) was added, and ethyl acetate (100 ml x 3) was used for extraction. The organic phases were combined and concentrated under reduced pressure. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column to obtain 70 mg of the product.

5. Synthesis of C89

In a 100 mL sealed bottle, C89-5 (45 mg, 1.0 eq), palladium acetate (10.42 mg, 0.2 eq), XantPHos (50 mg, 0.4 eq), t-BuOK (48.3 mg, 2 eq), int1E (97.4 mg, 1.2 eq) were added. Dioxane (8 ml) was added and reacted at 100°C for 2 h. After the reaction was completed, the mixture was filtered, concentrated, and purified by high performance liquid chromatography to obtain 1.2 mg of the product.

Embodiment 21

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C90-1

Add int2G (80 mg, 1.00 eq), int1E (105 mg, 1.00 eq), cesium carbonate (272 mg, 3.00 eq), XantpHos (32 mg, 0.20 eq), palladium acetate (6.2 mg, 0.10 eq) and 1,4-dioxane (5 mL) to a glass tube and replace the nitrogen. Heat to 100 °C and react for 2 h. The reaction is complete. Cool the reaction solution, filter, wash the solid with ethyl acetate, and concentrate the filtrate under reduced pressure. Purify the product by TLC preparative plate to obtain 30 mg. Ms[M+H] ⁺ : 519.3.

2. Synthesis of C90

Compound C90-1 (30 mg, 1.00 eq) was dissolved in ethyl acetate (3 mL), and a solution of hydrogen chloride in dioxane was added under stirring and stirred at room temperature for 20 minutes. After the reaction was complete, ethyl acetate and water were added for extraction, and the aqueous phase was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Purification by thin layer chromatography preparative plate gave 31 mg of the product. ¹ H NMR (400MHz, DMSO-d6) δppm 12.03(s,1H),8.27(s,1H),7.85(s,1H),7.76(s,1H),5.92(s,1H),4.99(s,1H),3.18(s,3H),3.07-3.03(m,1H),2.35(s, 1H),2.04-2.02(m,1H),1.90(m,8H),1.72(m,2H),1.64-1.61(m,3H),1.53-1.50(m,2H).Ms[M+H] ⁺ : 483.1.

Embodiment 22

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C91-2

N,N-dimethylformamide (20 mL) was added to a 100 mL flask, NaH (2.25 g, 5.0 eq) was added at 0°C, dimethyl malonate (9 g, 5.0 eq) was added at room temperature, and the mixture was reacted at 40°C for 40 min, then C91-1 was added and the temperature was raised to 100°C for further reaction for 16 h. After the reaction was completed, N,N-dimethylformamide was removed by concentration, water (20 mL) was added, and impurities were removed by extraction with ethyl acetate (4*20 mL), the aqueous phase was retained, concentrated H ₂ SO ₄ was added thereto to adjust the pH until a large amount of solid precipitated, the solid was collected, and washed with water several times, and finally dried to obtain 2.5 g of crude product, which was directly used in the next step.

2. Synthesis of C91-3

In a 100 mL flask, add crude C91-2 (2.3 g, 1.0 eq), concentrated HCl (22 mL) and AcOH (16 mL), react at 100 °C for 6 h. After the reaction is complete, concentrate to obtain 2.5 g of a crude product, which is directly used in the next step.

3. Synthesis of C91-4

C91-3 (1.5 g, 1.0 eq) was added to a 50 mL flask, dissolved in anhydrous dichloromethane (20 mL) and stirred for 3-5 min. Zn(OTf) ₂ (0.6 g, 0.2 eq) and DBU (2.5 g, 2.0 eq) were added in sequence at room temperature, and finally diphenyl (vinyl) sulfonium trifluoromethanesulfonate (1.1 g, 0.4 eq) was added, and the reaction was carried out at room temperature for 15 min. After the reaction was completed, water was added to the reaction solution, and the dichloromethane was extracted three times. The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 180 mg of the product.

4. Synthesis of C91-5

C91-4 (100 mg, 1.0 eq) was added to a 50 mL flask, dissolved in anhydrous N, N-dimethylformamide (5 mL), and NaH (40 mg, 2.0 eq) was added at 0°C. The mixture was reacted at 0°C for 0.5 h, and then CH ₃ I (100 mg, 1.5 eq) was added. The reaction was continued for 1.5 h. After the reaction was completed, water was added to the reaction solution, and then extracted with ethyl acetate three times. The organic phases were combined, washed with saturated aqueous ammonium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified to obtain 110 mg of the product.

5. Synthesis of C91-6

C91-5 (50 mg, 1.0 eq), int1E (75 mg, 1.0 eq) and DMSO (3 mL) were added to a 25 mL flask as solvent, and potassium tert-butoxide (63 mg, 2.5 eq) was added thereto under stirring at room temperature, and the mixture was reacted at room temperature for 20 min. After the reaction was completed, water was added to the reaction solution, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with saturated aqueous ammonium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified to obtain 17 mg of the product.

7. Synthesis of C91

C91-6 (17 mg, 1.0 eq) and ethyl acetate (3 mL) as solvent were added to a 25 mL flask, and dioxane hydrochloride solution (1 M, 1 mL) was added thereto under stirring at room temperature, and the reaction was carried out at room temperature for 10 min. After the reaction was completed, the reaction solution was added to a saturated sodium bicarbonate aqueous solution in an ice bath, and extracted with ethyl acetate three times. The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified to obtain 8 mg of the product. Ms[M+H] ⁺ : 464.5.

Embodiment 23

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C95-2

Dissolve compound C95-1 (5.0 g, 42.37 mmol) in N,N-dimethylformamide (40 mL), cool to 0°C, add sodium hydride (2.03 g, 50.85 mmol), stir at 0°C for 15 minutes, then drop iodomethane (7.2 g, 50.85 mmol), slowly warm to room temperature after dropwise addition, and stir for 3 hours. The reaction is complete. Cool, add appropriate amount of water to quench the reaction, concentrate under reduced pressure, and purify by column chromatography to obtain 5.6 g of crude product, Ms[M+H] ⁺ : 133.1.

2. Synthesis of C95-3

Compound C95-2 (5.6 g, 42.4 mmol) was dissolved in tert-butyl alcohol (50 mL), sodium bicarbonate (10.7 g, 127.27 mmol) was added, stirred for 10 minutes, cooled to 0°C, and then bromine (40.7 g, 254.55 mmol) was added dropwise. The temperature was slowly raised to room temperature after the addition, and stirred overnight. The temperature was lowered, and an appropriate amount of water was added to quench the reaction. The organic phase was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 1.81 g of the product, Ms[M+H] ⁺ : 382.8.

3. Synthesis of C95-4

Dissolve compound C95-3 (1.7 g, 4.45 mmol) in acetic acid solution (25 mL), cool to 0°C, add zinc powder (1.46 g, 22.25 mmol) in batches, warm to room temperature and stir for 15 minutes. The reaction is complete. Filter, concentrate under reduced pressure, and purify by column chromatography to obtain 0.99 g of the product, Ms[M+H] ⁺ : 226.0.

4. Synthesis of C95-5

Compound C95-4 (0.95 g, 4.20 mmol) was dissolved in dichloromethane solution (25 mL), diphenyl (vinyl) sulfonium trifluoromethanesulfonate (1.52 g, 4.20 mmol), zinc trifluoromethanesulfonate (305 mg, 0.84 mmol) were added, stirred at room temperature for 3 minutes, 1,8-diazobispiro[5.4.0]undec-7-ene (1.28 g, 8.40 mmol) was added, and stirred at room temperature for 10 minutes. The reaction was complete. The reaction solution was diluted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 667 mg of the product, Ms[M+H] ⁺ : 253.0.

5. Synthesis of C95

Compound C95-5 (100 mg, 0.40 mmol), int1E (120 mg, 0.32 mmol), palladium acetate (18 mg, 0.08mmol), XantpHos (68mg, 12mmol), cesium carbonate (388mg, 1.19mmol) and 1,4-dioxane (15mL), heated to 100℃ under nitrogen protection for 12 hours. After the reaction was completed, filtered, concentrated, and purified by TLC preparation plate to obtain 9mg of the product. ¹ H NMR (400MHz, DMSO-d ₆ )δ11.55(s,1H),8.20(s,1H),7.87(s,1H),7.80-7.68(m,1H),7.49(s,1H),5.42(s,1H),4.99-4.93(m,1H),3.29(s,3H),3.04-2.94(m,1H),2.45- 2.38(m,1H),2.36-2.31(m,1H),2.01-1.95(m,1H),1.92-1.79(m,8H),1.75-1.72(m,2H),1.71-1.61(m,2H),1.59-1.56(m,2H).Ms[M+H] ⁺ : 449.3.

Embodiment 24

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C59

In a 100 mL single-mouth bottle, int1E (150 mg 1 eq), int2A (80 mg 1 eq), and DMSO (5 ml) were added and stirred to dissolve, and then potassium tert-butoxide (133.68 mg 3 eq) was added and stirred at room temperature for 20 min. After the reaction was completed, saturated sodium chloride (20 ml) was added to the reaction solution, and ethyl acetate (50 ml x ³ ) was extracted. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified to obtain 40 mg of the product. NMR (400MHz, DMSO) δ＝12.11(s,1H),8.54(s,1H),8.31(s,1H),7.78(s,1H),6.26(s,1H),5.01(s,1H),3.50(d,J＝14.7,3H),3.08(d,J＝8.1,1H),2.46(d,J ＝7.9,1H),2.35(s,1H),2.04(s,1H),1.91(s,7H),1.82–1.69(m,4H),1.65(dd,J＝7.8,3.7,3H).Ms[M+H] ⁺ : 450.2.

Embodiment 25

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C94

In a 100 mL single-mouth bottle, int29 (150 mg 1 eq), int2A (74 mg 1 eq), and DMSO (5 ml) were added and stirred to dissolve, and then potassium tert-butoxide (120 mg 3 eq) was added and stirred at room temperature for 20 min. After the reaction was completed, saturated sodium chloride 20 ml was added to the reaction solution, and ethyl acetate (50 ml x 3) was used for extraction. The organic phase was dried over anhydrous ^sodium sulfate, filtered, concentrated, and purified to obtain 27 mg of the product. NMR (400MHz, DMSO-d6) δ12.10(s,1H),8.53(s,1H),8.30(s,1H),7.21(d,J＝8.4Hz,1H),6.24(s,1H),5.02(s,1H),3.97–3.75(m,1H),3.47(s,3H),3.14–3 .01(m,1H),2.43–2.25(m,2H),2.10–1.98(m,1H),1.92(s,1H),1.81–1.69(m,4H),1.65(q,J＝4.2,3.6Hz,3H),1.13(d,J＝6.8Hz,3H).Ms[M+H] ⁺ ：494 .2.

Embodiment 26

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C103-1

Compounds int31 (130 mg, 1.0 eq) and int2A (74.1 mg, 1.0 eq) were dissolved in dimethyl sulfoxide (13 mL), and a dimethyl sulfoxide solution of potassium tert-butoxide (78 mg, 2.0 eq) was slowly added dropwise under an ice-water bath. After addition, the mixture was reacted at room temperature for 20 minutes. Purified water was added dropwise under an ice-water bath, and the mixture was extracted with ethyl acetate. The organic phase was then washed with a saturated sodium chloride solution, concentrated in vacuo until no liquid flowed out, and 100 mg of the product was prepared by thin-layer chromatography. Ms[M+H] ⁺ : 540.2.

2. Synthesis of C103

Compound C103-1 (100 mg, 1.0 eq) was dissolved in 10 mL of ethyl acetate, and a 4 mol/L hydrogen chloride solution in 1,4-dioxane (2 mL) was slowly added dropwise under an ice-water bath. After addition, the mixture was reacted at room temperature for 10 minutes. The reaction system was added dropwise to a saturated sodium bicarbonate solution under an ice bath, and the liquid was extracted with ethyl acetate. The organic phase was then washed with a saturated sodium chloride solution, concentrated in vacuo until no liquid flowed out, and 7.39 mg of the product was prepared by thin layer chromatography, ¹ H NMR (400 MHz, DMSO-d6) δppm 12.09(s,1H),8.52(s,1H),8.29(s,1H),6.89(d,J＝8.4Hz,1H),6.24(s,1H),4.99(s,1H),3.46(s,3H),3.07(t,J＝8.8Hz,1H),2.67(s,1H),2.33(s, 1H),2.09–1.98(m,1H),1.90(s,1H),1.82–1.56(m,6H),1.43–1.17(m,3H),1.00(d,J＝6.8Hz,3H),0.79(t,J＝7.2Hz,3H).Ms[M+H] ⁺ : 440.2.

Embodiment 27

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C104

In a 50 mL single-mouth bottle, int32 (146 mg, 1 eq), int2A (74.1 mg, 1.0 eq) and DMSO (5 ml) were added and stirred to dissolve, and then potassium tert-butoxide (120 mg 3 eq) was added and stirred at room temperature for 20 min. After the reaction was completed, saturated sodium chloride (20 ml) was added to the reaction solution, and ethyl acetate (50 ml x 3) was extracted. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and separated by high performance liquid chromatography to obtain ³⁵ mg of the product. NMR(400MHz,Chloroform-d)δ8.44(s,1H),7.19(s,1H),6.54(s,1H),5.19(q,J＝5.0,4.6Hz,1H),4.91(s,1H),3.65(s,3H),3.21(p,J＝8.2Hz,1H),2.54(s,1H), 2.29(s,2H),2.14(dd,J＝11.6,6.3Hz,1H),1.99–1.75(m,13H),1.42(s,3H).Ms[M+H] ⁺ : 452.5.

Embodiment 28

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C106-1

Compound int2F (800 mg, 1.00 eq), N-bromosuccinimide (3.85 g, 4.00 eq) and carbon tetrachloride (30 mL) were added to a 100 mL glass flask, and azobisisobutyronitrile (59 mg, 0.10 eq) was added at room temperature. The reaction solution was reacted at 75 °C for 16 h in a nitrogen atmosphere. The reaction solution was cooled to room temperature, concentrated, and purified by silica gel column to obtain 600 mg of the product. Ms[M+H] ⁺ : 378.9.

2. Synthesis of C106-2

C106-1 (400 mg, 1.00 eq) and ethanol (12 mL) were added to a 50 mL single-mouth bottle. Silver nitrate (630 mg, 3.50 eq) in water (8 mL) was added at room temperature. The reaction solution was stirred at 80°C for 1 h. Filtered and the liquid phase was concentrated. The aqueous phase was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the organic phase was concentrated. The product was purified by TLC preparative plate to obtain 190 mg. Ms[M+H] ⁺ : 237.0.

3. Synthesis of C106-3

Add C106-2 (190 mg, 1.00 eq) and dichloromethane (20 mL) to a 50 mL three-necked flask. Cool to -50 °C and slowly add diethylaminosulfur trifluoride (518 mg, 4.00 eq). Stir the reaction solution at -50 °C for 10 min, then slowly warm to room temperature and react for 2 h. Saturated sodium bicarbonate aqueous solution was slowly added at 0 °C, and the aqueous phase was extracted with dichloromethane. Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate. Purify the product on a thin layer chromatography preparative plate to obtain 170 mg. Ms[M+H] ⁺ : 259.0.

4. Synthesis of C106-4

C106-3 (80 mg, 1.00 eq), int7 (103 mg, 1.00 eq), cesium carbonate (303 mg, 3.00 eq), XantpHos (36 mg, 0.20 eq), palladium acetate (7 mg, 0.10 eq) and 1,4-dioxane (5 mL) were added to a 25 mL glass tube. The mixture was heated to 100 °C in a nitrogen atmosphere and reacted for 2 h. The reaction solution was cooled, filtered, the residue was washed with ethyl acetate, and the organic phase was concentrated under reduced pressure. The product was purified by TLC preparative plate to obtain 100 mg. Ms[M+H] ⁺ : 555.3.

5. Synthesis of C106

Compound C106-4 (90 mg, 1.00 eq) was dissolved in formic acid (4 mL), heated to 80°C for 1.5 hours, and the reaction solution was cooled and concentrated. Ethyl acetate was dissolved, and the pH was adjusted to alkaline with saturated sodium bicarbonate aqueous solution. The aqueous phase was extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and concentrated. 14 mg of the product was prepared. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 12.10 (s, 1H), 8.09 (s, 1H), 7.83-7.69 (m, 2H), 6.76 (t, J=55.2 Hz, 1H), 5.81 (s, 1H), 5.00 (s, 1H), 3.51 (s, 3H), 3.13-3.05 (m, 1H), 2.50-2.43 (m, 2H), 2.39-2.30 (m, 1H), 2.09-2.02 (m, 1H), 1.99-1.79 (m, 7H), 1.79-1.65 (m, 4H), 1.63-1.59 (m, 2H). Ms [M+H] ⁺ : 499.2.

Embodiment 29

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C107-1

Add compound int2B (500 mg, 1.00 eq) and tetrahydrofuran (15 mL) to a 50 mL glass flask, cool to 0°C and add lithium aluminum hydride (110 mg, 1.20 eq). Heat the reaction solution to 60°C and stir for 1 h. Cool the reaction solution to 0°C, quench with ice water, filter, wash the filter residue with ethyl acetate, extract the aqueous phase with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, and concentrate. Purify with a silica gel column to obtain 150 mg of the product. Ms[M+H] ⁺ : 195.1.

2. Synthesis of C107-2

C107-1 (100 mg, 1.00 eq), int7 (171 mg, 1.00 eq), cesium carbonate (504 mg, 3.00 eq), XantpHos (60 mg, 0.20 eq), palladium acetate (12 mg, 0.10 eq) and 1,4-dioxane (5 mL) were added to a 25 mL glass tube. The mixture was heated to 100 °C in a nitrogen atmosphere and reacted for 2 h. The mixture was cooled, filtered, and the organic phase was concentrated. The product was purified by TLC preparative plate to obtain 150 mg. Ms [M+H] ⁺ : 491.3.

3. Synthesis of C107

Compound C107-2 (130 mg, 1.00 eq) was dissolved in formic acid (4 mL), heated to 80°C for 1.5 hours, cooled, concentrated, dissolved in ethyl acetate, adjusted pH to alkaline with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, dried the organic phase with anhydrous sodium sulfate, and concentrated. 16 mg of the product was obtained, ¹ H NMR (400 MHz, DMSO-d6) δ ppm 11.87(s,1H),7.81-7.66(m,2H),7.61(d,J＝5.6Hz,1H),7.37(d,J＝5.6Hz,1H),5.76(s,1H),4.99(s,1H),3.38(s,2H),3.11-2.96(m,1H),2.77(s,3 H),2.49-2.42(m,2H),2.38-2.30(m,1H),2.06-1.97(m,1H),1.89(s,6H),1.78-1.64(m,2H),1.63-1.52(m,1H),1.01(q,J＝3.6Hz,2H),0.88(q,J＝ 3.6Hz,2H).Ms[M+H] ⁺ :435.2.

Embodiment 30

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C109-2

Compound C109-1 (2.00 g, 1.00 eq), sodium bicarbonate (3.11 g, 3.00 eq) and tert-butyl alcohol (40 mL) were added to a 100 mL single-mouth bottle and stirred at room temperature for 10 min. Bromine (11.85 g, 6.00 eq) was slowly added dropwise. The reaction was allowed to react at room temperature for 1 h, and water was added to quench the reaction. The aqueous phase was extracted with ethyl acetate and separated. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by silica gel column gave 4.40 g of the product. Ms[M+H] ⁺ : 412.8.

2. Synthesis of C109-3

C109-2 (4.40 g, 1.00 eq) and methanol (80 mL) were added to a 250 mL single-mouth bottle. 10% Pd/C (4.00 g) was added at room temperature to replace the hydrogen. The reaction solution was stirred at 50°C for 24 h in a hydrogen atmosphere, filtered, and the filter cake was washed with methanol. The organic phase was concentrated to obtain 3.20 g of crude product. Ms[M+H] ⁺ : 179.1.

3. Synthesis of C109-4

C109-3 (3.20 g, crude product), diphenyl (vinyl) sulfonium trifluoromethanesulfonate (3.86 g, 1.00 eq) and zinc trifluoromethanesulfonate (775 mg, 0.20 eq) were dissolved in dichloromethane (40 mL), stirred at room temperature for 3 min, and 1,8-diazabicyclo [5.4.0] undec-7-ene (3.25 g, 2.00 eq) was slowly added dropwise to the system. The reaction was carried out at room temperature for 10 min, and a saturated aqueous solution of ammonium chloride was added to the reaction solution. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by silica gel column gave 900 mg of the product. Ms [M + H] ⁺ : 205.1.

4. Synthesis of C109-5

C109-4 (550 mg, 1.00 eq), sodium iodide (809 mg, 2.00 eq) and acetonitrile (15 mL) were added to a 50 mL single-mouth bottle. Trimethylsilyl chloride (588 mg, 2.00 eq) was added at room temperature to replace the hydrogen. The reaction solution was stirred at 50 ° C for 16 h in a hydrogen atmosphere, and the pH was adjusted to neutral with saturated sodium bicarbonate aqueous solution, filtered, and the liquid phase was concentrated. Purification on a silica gel column gave 700 mg of the product. Ms [M + H] ⁺ : 191.1.

5. Synthesis of C109-6

C109-5 (300 mg, 1.00 eq), triethylamine (957 mg, 6.00 eq) and dichloromethane (15 mL) were added to a 50 mL single-mouth bottle. Trifluoromethanesulfonic anhydride (668 mg, 1.50 eq) was dissolved in dichloromethane (5 mL) and slowly added dropwise to the reaction system under an ice bath. The reaction solution was stirred under an ice bath for 5 min, heated to room temperature and stirred for 30 min, saturated sodium bicarbonate aqueous solution was added to the reaction solution, the aqueous phase was extracted with dichloromethane, the organic phase was dried over anhydrous sodium sulfate and concentrated. The product was purified by TLC preparative plate to obtain 147 mg. Ms[M+H] ⁺ : 323.0.

6. Synthesis of C109-7

C109-6 (110 mg, 1.00 eq), int7 (113 mg, 1.00 eq), cesium carbonate (334 mg, 3.00 eq), XantpHos (14 mg, 0.20 eq), palladium acetate (3 mg, 0.10 eq) and 1,4-dioxane (5 mL) were added to a glass tube and the nitrogen atmosphere was replaced. The temperature was raised to 100°C for reaction for 1 h, the temperature was lowered, the mixture was filtered, the solid was washed with ethyl acetate, and the mixture was concentrated under reduced pressure. The product was purified by TLC preparative plate to obtain 90 mg. Ms[M+H] ⁺ : 505.3.

7. Synthesis of C109

Compound C109-7 (90 mg, 1.00 eq) was dissolved in formic acid (5 mL), heated to 80°C and stirred for 40 min, cooled to room temperature, and concentrated under reduced pressure. Re-dissolved in ethyl acetate, the pH was adjusted to alkaline with saturated sodium bicarbonate aqueous solution, and the aqueous phase was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by TLC preparative plate to obtain 11 mg of the product, ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.98(d,J＝5.2Hz,1H),6.84(s,1H),6.45(d,J＝5.2Hz,1H),6.06(s,1H),5.28(s,1H),5.22-5.14(m,1H),3.62(s,3H),3.17(p,J＝8.4Hz,1H),2.56-2.4 4(m,1H),2.40(s,1H),2.16-2.07(m,1H),2.02(s,6H),1.96-1.90(m,2H),1.90-1.81(m,4H),1.62(q,J＝4.0Hz,2H).Ms[M+H] ⁺ 449.2.

Embodiment 31

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C120-1

In a 100 mL single-necked bottle, compounds int40 (620 mg, 1.00 eq) and int2A (439 mg, 1.30 eq) were dissolved in DMSO (30 mL). To the above solution was added a solution of potassium tert-butoxide (543 mg, 3.00 eq) in DMSO (3 mL) under water bath conditions. After the addition was complete, the mixture was reacted at room temperature for 1 hour. Water was added to the reaction solution, and the mixture was extracted twice with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 780 mg of a crude product.

Ms[M+H] ⁺ ：558.6

2. Synthesis of C120

In a 100 mL single-mouth bottle, compound C120-1 (780 mg, 1.00 eq) was dissolved in dichloromethane (30 mL). Trifluoroacetic acid (1 mL) was added dropwise to the above solution under water bath conditions. After addition, the mixture was reacted at room temperature for 1 hour. Saturated sodium bicarbonate aqueous solution was added to the reaction solution, and the mixture was extracted twice with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 415 mg of a crude product. The product was then beaten twice with ethyl acetate (30 mL) to obtain 275 mg of the product. ¹ H NMR (400MHz, DMSO-d ₆ ) δ12.10 (s, 1H), 8.50 (s, 1H), 8.30 (s, 1H), 7.01 (d, J = 8.4Hz, 1H), 6.26 (s, 1H), 5.01 (s, 1H), 4.48 (t, J = 5.9Hz, 1H), 4.36 (t, J = 5.9 Hz,1H),3.63(q,J＝7.0Hz,1H),3.47(s,3H),3.16–3.01(m,1H),2.50(s,2H),2.10–1.85(m,2H),1.83–1.53(m,8H),1.07(d,J＝6.7Hz,3H), Ms[M+H] ⁺ : 458.5.

Embodiment 32

Compounds of the present invention

The synthetic route and experimental process are as follows:

1. Synthesis of C128-2

Compound int1D (700 mg, 1.00 eq), diisopropylethylamine (822 mg, 5.00 eq) and tetrahydrofuran (6 mL) were added to a 25 mL glass sealed tube, and C128-1 (248 mg, 1.40 eq) was added at room temperature. The reaction solution was reacted at 55°C for 16 h, and the reaction solution was cooled to room temperature. A large amount of ethyl acetate was added to the reaction solution, and the organic phase was washed four times with 1N sodium hydroxide aqueous solution. The organic phase was washed with saturated brine and dried with anhydrous sodium sulfate. After purification on a silica gel column, 630 mg of the product was obtained, Ms[M+H] ⁺ : 515.3.

2. Synthesis of C128-3

C128-2 (300 mg, 1.00 eq) and methanol (15 mL) were added to a 100 mL single-mouth bottle. 10% Pd/C (80 mg) was added at room temperature to replace the hydrogen. The reaction solution was stirred at room temperature for 1.5 h in a hydrogen atmosphere, filtered, and the filter cake was washed with methanol. The organic phase was concentrated to obtain 230 mg of a crude product. Ms [M+H] ⁺ : 381.2.

3. Synthesis of C128-4

C128-3 (230 mg, 1.00 eq), int2A (127 mg, 1.00 eq) and dimethyl sulfoxide (5 mL) were added to a 25 mL glass flask. Potassium tert-butoxide (204 mg, 3.00 eq) was dissolved in DMSO (1 mL) and slowly added dropwise to the reaction system at room temperature. The reaction was stirred at room temperature for 30 min, water was added to the reaction solution, the aqueous phase was extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, and concentrated. The product was purified by TLC preparative plate to obtain 170 mg. Ms[M+H] ⁺ : 554.3.

4. Synthesis of C128-5

Compound C128-4 (170 mg, 1.00 eq) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (1 mL) was added to the reaction solution. The reaction solution was stirred at room temperature for 30 min, and the pH of the reaction solution was adjusted to neutral with an aqueous solution of sodium bicarbonate. The aqueous phase was extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The organic phase was purified by TLC preparative plate, and then slurried with ethyl acetate/petroleum ether and filtered to obtain 80 mg of the product. Ms[M+H] ⁺ : 470.2.

5. Synthesis of C128 and C129

Compound C128-5 (80 mg) was prepared and separated into:

C129(47mg) ¹ H NMR(400MHz,DMSO)δppm 12.11(s,1H),8.52(s,1H),8.30(s,1H),7.43(s,1H),6.27(s,1H),5.08- 4.85(m,2H),3.47(s,3H),3.16-3.00(m,1H),2.50-2.48(m,1H),2.47-2.31(m,4H),2.08-1.99(m,1H), 1.96-1.85(m,1H),1.81-1.69(m,4H),1.69-1.58(m,3H),1.25(s,3H).Ms[M+H] ⁺ : 470.2.

C128(16mg) ¹ H NMR(400MHz,DMSO)δppm 12.10(s,1H),8.52(s,1H),8.30(s,1H),7.31(s,1H),6.26(s,1H),5.17- 4.96(m,2H),3.47(s,3H),3.15-3.02 (m,1H),2.74-2.60(m,2H),2.50-2.48(m,1H),2.14-1.98(m,3H),1.97-1.84(m,1H),1.81-1.57(m,7H) ,1.38(s,3H).Ms[M+H] ⁺ : 470.2.

The following compounds were synthesized using the above synthesis method:

Cell proliferation inhibition assay

1. Experimental Materials

Comparative compound Palbociclib

2. Experimental plan

1. Cell plating

a) Preparation of cell suspension

b) removing the culture medium from the culture flask;

c) Rinse the cells once with PBS;

d) digest with trypsin and collect by centrifugation;

e) Resuspend in culture medium, count and adjust to appropriate concentration.

f) Add the cell suspension to each well of a 96-well plate at a volume of 80 μL, with one plate for each cell type. 37°C, Incubate overnight in a 5% CO ₂ incubator.

2. Compound treatment

Compound dilution

a) Preparation of gradient dilution solutions of the test compounds: Palbociclib and the example compounds were prepared at 10 mM as stock solutions. Then 2.5 μL of the stock solution was dissolved in 497.5 μL of DMSO-free culture medium, and then 3-fold continuous gradient dilutions were performed with 0.1% DMSO culture medium, for a total of 9 concentrations. The compound concentrations after dilution were as follows:

10000nM, 3333.33nM, 1111.11nM, 370.34nM, 123.45nM, 41.15nM, 13.72nM, 4.57nM, 1.52nM

b) After thorough mixing, 20 μL of the culture compound solution was added to the cell culture plate containing 80 μL of cells, with 4 replicate wells for each concentration.

c) Transfer the cells to an incubator and incubate for 5 days.

3. MTT assay

a) Take out the cell culture plate and add 10μL of 5mg/ml MTT into the hood.

b) Place the cell culture plate back into the incubator and continue incubating for 3 hours

c) Take out the cell culture plate and remove the culture medium, add 100uL of isopropanol (0.4mM HCl, 0.4% NP40), shake at room temperature for 30 minutes, and read the plate at 570nm.

4. Data Analysis

The survival rate (%Cell Survival) is calculated using the following formula:

%Cell Survival＝100%×(OD_Sample-OD_LCave)/(OD_HC-OD_LCave)

OD_HC: 0.1‰ DMSO control group cell reading

OD_Sample: Cell reading after adding compound

OD_LC: Blank culture medium reading

Analyzed by Prizm:Dose-response-Inhibition-Log(inhibitor) vs response(three parameters for the best fit).

The results of the cell proliferation inhibition test are shown in Table 1.

Table 1

A≤300nM; 300nM<B≤1000nM; 1000nM<C≤3000nM; D>3000nM

The experimental results showed that for the CyclinE-amplified cell line OVCAR3, the example compound It showed good inhibitory effect, while for the non-CyclinE amplified cell line TOV21G, the inhibitory effect of the example compound was not obvious, showing the good selectivity of the compound.

Enzyme activity test

Prepare the compound:

1. Prepare the test compound and palbociclib into 0.5 nM DMSO solution;

2. Use Echo550 to transfer 20 μL of the stock solution to a 384-well plate. Use DMSO as a blank control.

Experimental steps:

1. Prepare 1.3X enzyme solution containing enzyme, substrate, and coenzyme factor according to the table below;

2. Add 15uL of 1.3x enzyme solution to each well and incubate at room temperature for 30 minutes;

3. Add 5 μL of 4x ATP solution to start the reaction. The final volume of each well is 20 μL;

4. After incubation at room temperature for 150 min, add 75 μL of stop buffer solution to terminate the experiment;

5. Analyze samples using EZ reader.

Data Analysis:

Calculate the % inhibition using the following equation

The positive control (PC) was treated with DMSO

The negative control (NC) was without enzyme.

% inhibition rate = 100-100*((CR _PC -CR _sample )/(CR _PC -CR _NC ))

Table 2

A≤10nM; 10nM<B≤50nM; 50nM<C≤500nM; D>500nM

The experimental results show that the compounds of the present invention have excellent selectivity and activity for CDK2 kinase.

The present invention provides a series of compounds with excellent CDK2 kinase activity and selectivity. The series of compounds have good inhibitory effects on excessive cell proliferation caused by abnormal expression or amplification of CyclinE and have broad application prospects.

All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims (10)

A compound, characterized in that the compound is a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof,
in: Ring A is a saturated or partially saturated 5-6 membered heterocycloalkyl group containing 1 or 2 heteroatoms selected from N, O or S; Ring B is selected from the group consisting of C _5-6 aryl, 5-6 membered heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; Each _Ra , _Rc , and _Rd is independently selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, cyano, =O, _C1-4 alkyl, C1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated _C1-4 alkyl, deuterated _C1-4 alkyl, halogenated _C1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, _C1-4 alkoxy, halogenated _C1-4 alkoxy, and _C3-4 cycloalkyl _; is a single bond or a double bond; p, q, t, w are each independently selected from the following group: 0, 1, 2, 3, 4; s is selected from the following group: 1, 2; R ₁ is selected from the following group: -CONR ₂ R ₃ , -COOR ₂ , 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is optionally substituted with 0, 1, 2, 3, 4, 5 or 6 R ₄ ; _X1 is selected from the group consisting of _CH2 , O; or _X1 is absent, such that Four-membered cyclobutyl X ₂ is selected from the group consisting of O, -CH ₂ O-, NH, -CH ₂ NH-; R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _3-12 cycloalkyl), -L-( _3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, amino, C _1-4 alkyl, C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl, Halogenated C _1-4 alkoxy, -S(O) ₂ -C _1-4 alkyl, Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy; L is a bond or a C _1-6 alkylene group, wherein the C _1-6 alkylene group is optionally substituted by 0, 1, 2 or 3 substituents selected from the group consisting of deuterium, =O, hydroxyl, halogen, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo-C _1-4 alkyl, halo-C _1-4 alkoxy; Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy; R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl; Each R ₇ is independently selected from the group consisting of hydrogen, deuterium, halogen, C _1-4 alkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl. The compound according to claim 1, characterized in that the compound has a structure selected from the group consisting of:
Among them, ring A, ring B, R ₁ , _Ra , R _c , R _d , p, q, t are as defined in claim 1. The compound according to claim 2, characterized in that the compound has a structure selected from the group consisting of:
Among them, ring A, ring B, _Ra , _Rc , _Rd , p, q, t as defined in claim 1; R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _{3-12 cycloalkyl} ), -L-(3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, _{cyano, =O, C 1-4 alkyl} , C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl _{, halogenated C 1-4 alkoxy} , Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy; L as defined in claim 1; Ring C is selected from the following group: 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ; Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl C _1-6 alkyl, C _1-6 alkoxy, C _{1-6 alkoxy halogenated C 1-6 alkyl, C 1-6 alkoxy} halogenated C _1-6 alkoxy halogenated C _1-4 alkyl, C _1-4 alkoxy halogenated ... R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl. The compound according to claim 3, characterized in that the compound has a structure selected from the group consisting of:
Among them, ring A, ring B, _Ra , _Rc , _Rd , p, q, t as defined in claim 1; R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _3- wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted by 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, C _1-4 alkyl, C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl _, halogenated C _1-4 alkyl, halogenated _{C 1-4 alkoxy} , Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy; L as defined in claim 1; Ring C is selected from the following group: 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ; Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy; R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl. The compound according to claim 1, characterized in that the compound has a structure selected from the group consisting of:

in, R ₁ is selected from the following group: -CONR ₂ R ₃ , 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ; R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, deuterated C _1-6 alkyl, -L-(C _{3-12 cycloalkyl} ), -L-(3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S), wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, _{cyano, =O, C 1-4 alkyl} , C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl _{, halogenated C 1-4 alkoxy} , Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy; L as defined in claim 1; Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy; R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl; R _a1 is selected from the group consisting of hydrogen, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, deuterated C 1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, ₂ or 3 heteroatoms selected from N, O or S, and C _3-4 cycloalkyl; Each R _c1 is independently selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, cyano, =O, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-4 alkoxy, halogenated C _1-4 alkoxy; Each R _d1 is independently selected from the group consisting of hydrogen, deuterium, halogen, cyano, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-4 alkoxy, halogenated C _1-4 alkoxy; m and r are each independently selected from the following group: 0, 1, 2, 3. The compound according to claim 5, characterized in that the compound has a structure selected from the group consisting of:
in, R ₁ is selected from the following group: -CONR ₂ R ₃ , 3-6 membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 5-6 membered monocyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, 7-10 membered bicyclic heteroaryl containing 1, 2, 3 or 4 heteroatoms selected from N, O or S, wherein the 3-6 membered heterocycloalkyl, 5-6 membered monocyclic heteroaryl, 7-10 membered bicyclic heteroaryl is substituted by 0, 1, 2, 3, 4, 5 or 6 R ₄ ; _R2 and _R3 are each independently selected from the following group: hydrogen, _C1-6 alkyl, deuterated _C1-6 alkyl, -L-( _C3-12 cycloalkyl), -L-( _3-12 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S); wherein the C _1-6 alkyl, C _3-12 cycloalkyl, 3-12 membered heterocycloalkyl is optionally substituted by 0, 1, 2, 3 or ₄ substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, C _1-4 alkyl, C _3-6 cycloalkyl, C _1-4 alkoxy, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl, halogenated C _{1-4 alkoxy} , Alternatively, _R2 and _R3 together with the N to which they are attached form a 3-7 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the 3-7 membered heterocycloalkyl group is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, hydroxyl, halogen, cyano, =O, _C1-4 alkyl, _C1-4 alkoxy, deuterated _C1-4 alkyl, halogenated _C1-4 alkyl, halogenated _C1-4 alkoxy; L as defined in claim 1; Each R ₄ is independently selected from the following group: deuterium, hydroxyl, halogen, cyano, =O, _{C 1-6} alkyl, C _3-6 cycloalkyl, deuterated C _1-6 alkyl, C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-6 alkoxy, halogenated C _1-6 alkoxy, -SO ₂ R ₅ , -SO ₂ (NR ₅ R ₆ ), -CO(OR ₅ ), -CO(NR ₅ R ₆ ), wherein the C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 heteroalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 heteroalkyl, C _1-6 alkoxy, halogenated C 1-6 _1-6 alkoxy is optionally substituted with 0, 1, 2, 3 or 4 substituents selected from the group consisting of deuterium, =0, halogen, hydroxy, cyano, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl, halo C _1-4 alkoxy; R ₅ and R ₆ are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-6 cycloalkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl; R _a1 is selected from the group consisting of hydrogen, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, deuterated C 1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, ₂ or 3 heteroatoms selected from N, O or S; Each R _c1 is independently selected from the following group: hydrogen, deuterium, halogen, hydroxyl, cyano, =O, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-4 alkoxy, halogenated C _1-4 alkoxy; Each R _d1 is independently selected from the group consisting of hydrogen, deuterium, halogen, cyano, C _1-4 alkyl, C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, halogenated C _1-4 alkyl, halogenated C _1-4 heteroalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C _1-4 alkoxy, halogenated C _1-4 alkoxy; m and r are each independently selected from the following group: 0, 1, 2, 3. The compound according to claim 1, characterized in that the compound is selected from the group consisting of:











A pharmaceutical composition comprising a preventive and/or therapeutically effective amount of or more compounds according to claim 1 and a pharmaceutically acceptable carrier. A use of the compound according to claim 1, characterized in that it is used to prepare a drug for regulating CDK activity or for preventing and/or treating CDK-related diseases. A use of the compound according to claim 1, characterized in that it is used to prepare a drug for preventing and/or treating cell cycle protein-related diseases.